Methods and treatment of trauma

Abstract

A method for reversing hemorrhagic shock or hemorrhagic trauma. [Selected Figure] Figure 1

Description

Technical Field

[0001] Cross - Reference to Related Applications This application claims priority to U.S. Provisional Application No. 62 / 508,783, filed on May 19, 2017, the entire disclosure of which is hereby incorporated by reference.

[0002] Government Rights This invention was made with government support under R44HL132172, awarded by the National Heart, Lung, and Blood Institute. The government has certain rights in this invention.

[0003] Field of the Invention This disclosure relates to the treatment of trauma and hemorrhagic shock.

Background Art

[0004] In 2010, there were 5.1 million deaths from injuries, exceeding the combined number of deaths from HIV, tuberculosis, and malaria (3.8 million). See Norton, et al., “Global Health Injuries” (The NEJM, 368:1723-30 (2013)) (“Norton 2013”) (this entire work is incorporated herein by reference). Injuries include unintentional injuries (e.g., traffic accidents, falls, and burns) and intentional injuries (e.g., self-injury, interpersonal violence, war, and conflict). See Norton 2013. Deaths from injuries increased 24% worldwide between 1990 and 2010, and 23% in the United States between 2000 and 2010. See Norton 2013. In addition, at least 20% of all trauma deaths result from viable injuries, and therefore such trauma deaths are preventable with optimal care. Fox et al., “Earlier Endpoints are Required for Hemorrhagic Shock Trials Among Severely Injured Patients.” Shock, 47:567-73 (2017) (this entire work is incorporated herein by reference). Given the percentage of preventable deaths, there is an urgent need to develop therapies for avoidable complications that lead to death.

[0005] Penetrating wounds (e.g., gunshot or stab wounds) and blunt trauma (e.g., injuries from falls or car accidents) are the primary causes of hemorrhagic trauma. The resulting shock is a state of insufficient oxygen supply to tissues due to massive bleeding, which leads to oxygen debt, anaerobic metabolism, and increased plasma lactate levels. If the shock cannot be reversed by the restoration of circulation and oxygen delivery, it can lead to permanent tissue damage, multiple organ failure, and death.

[0006] Clinical complications of hemorrhagic trauma and shock include death due to blood loss within hours of trauma, and death 24 hours after the onset of pathological conditions resulting from trauma and massive transfusion. These pathological conditions include multiple organ failure (including lung, kidney, and liver failure from acute traumatic coagulation disorder or inflammation), and infection / sepsis from transfusion-associated immunomodulation. All of these pathological conditions are exacerbated by lower quality blood products and larger quantities of pRBCs transfused.

[0007] One approach to treating hemorrhagic shock is the use of crystalloids for resuscitation. However, the use of crystalloids leads to an increased risk of pathological conditions and death by causing trauma-induced coagulation disorders. For at least this reason, early administration of blood components to reverse shock caused by hemorrhagic trauma is advocated. Concentrated red blood cells (pRBCs) are transfused to patients with hemorrhagic trauma to restore lost blood volume, restore oxygen-carrying capacity in the patient, and restore oxidative metabolism in tissues from anaerobic metabolism. However, the use of pRBCs is not without risks of complications, including antigen mismatch, pathogen transmission, circulatory overload, and degradation of pRBCs during ex vivo storage.

[0008] Stored blood, when stored conventionally, undergoes steady deterioration that leads to various storage lesions (particularly hemolysis, hemoglobin degradation, and a decrease in ATP and 2,3-DPG concentrations). The effects of this steady deterioration during storage manifest, for example, as a reduction in the 24-hour in vivo recovery rate when transfused to a patient. The rapid decrease in hematocrit resulting from the reduced 24-hour recovery rate can lead to delayed hemolytic transfusion reactions (DHTR) in severe cases. Other complications, such as systemic inflammatory response syndrome (SIRS), transfusion-associated acute lung injury (TRALI), and transfusion-associated immunomodulation (TRIM), are linked to transfusions of stored blood, but the underlying causes remain unclear.

[0009] Even when transfused within the current 6-week limit, stored RBCs exhibit lower quality (e.g., increased RBC fraction removal, reduced oxygen exchange capacity, decreased deformability) and increased toxicity, often manifesting as clinical complications of transfusion therapy. Numerous and growing literature supports this view. See: Zimring, “Established and theoretical factors to consider in assessing the red cell storage lesion,” Blood, 125:2185-90 (2015); Zhu et al., “Impaired adenosine-5'-triphosphate release from red blood cells promotes their adhesion to endothelial cells: a mechanism of hypoxemia after transfusion,” Critical care medicine,39:2478-86(2011);Weinberg et al.,“Red blood cell age and potentiation of transfusion-related pathology in trauma patients,”Transfusion,51:867-73(2011); Spinella et al.,“Does the storage duration of blood products affect outcomes in critically ill patients?”Transfusion 51:1644-50(2011);Roback et al.,“Insufficient nitric oxide bioavailability:a hypothesis to explain adverse effects of red blood cells transfusion,”Transfusion,51:859-66(2011);Reynolds et al.,“The transfusion problem:role of aberrant S-nitrosylation,”Transfusion,51:852-8(2011);Kim-Shapiro et al.,“Storage lesion:role of red blood cell breakdown,”Transfusion,51:844-51(2011);Jy et al.,“Microparticles in stored red blood cells as potential mediators of transfusion complications,”Transfusion,51:886-93(2011);Hod et al.,“Transfusion of human volunteers with older,stored red blood cells produces extravascular hemolysis and circulating non-transferrin-bound iron,”Blood,118:6675-82(2011); Flegel et al.,“Does prolonged storage of red blood cells cause harm?”British journal of haematology 165:3-16(2014);Redlin et al.,“Red blood cell storage duration is associated with various clinical outcomes in pediatric cardiac surgery,”Transfusion medicine and hemotherapy:offizielles Organ der Deutschen Gesellschaft fur Transfusionsmedizin und Immunhamatologie 41:146-51(2014);Rogers et al.,“Storage duration of red blood cell transfusion and Clostridium difficile infection:a within person comparison,”PLoS One 9:e89332(2014); Spinella et al.,“Properties of stored red blood cells:understanding immune and vascular reactivity,”Transfusion 51:894-900(2011);Brown et al.,“Length of red cell unit storage and risk for delirium after cardiac surgery,”Anesth Analg,119:242-50(2014);Wang et al.,“Transfusion of older stored blood worsens outcomes in canines depending on the presence and severity of pneumonia,”Transfusion,54:1712-24(2014);Liu et al.,“Mechanism of faster NO scavenging by older stored red blood cells,”Redox biology,2:211-9(2014);Prestia et al.,“Transfusion of stored blood impairs host defenses against Gram-negative pathogens in mice,”Transfusion 54:2842-51(2014);D’Alessandro et al.“An update on red blood cell storage lesions, as gleaned through biochemistry and omics technologies,” Transfusion, 55:205-19 (2015) (these entire works are incorporated herein by reference). Extensive in vitro studies have clearly demonstrated the degradation (storage alteration) of RBCs during conventional storage. A growing series of metabolome studies have indicated the occurrence of storage alteration at the molecular level. See: Roback et al., “Metabolomics of AS-1 RBCs Storage,” Transfusion medicine reviews (2014); D'Alessandro et al., “Metabolomics of AS-5 RBCs supernatants following routine storage,” Vox sanguinis (2014); D'Alessandro et al., “Routine storage of red blood cell (RBC) units in additive solution-3: a comprehensive investigation of the RBC metabolome,”Transfusion 55:1155-68(2015);D'Alessandro et al.,“Red blood cell storage in additive solution-7 preserves energy and redox metabolism: a metabolomics approach,”Transfusion(2015);Wither et al.,“Hemoglobin oxidation at functional amino acid residues during routine storage of red blood cells,”Transfusion(2015);D'Alessandro et al. al.,“Citrate metabolism in red blood cells stored in additive solution-3,” Transfusion (2016); D'Alessandro et al., “Omics markers of the red cell storage lesion and metabolic linkage,” Blood Transfus, 15:137-44 (2017) (these are incorporated herein by reference in their entirety). There is a need to reduce or prevent this degradation to increase the effectiveness of transfusions (delivering more O2 to peripheral tissues immediately after transfusion) and to reduce deaths from hemorrhagic trauma.

[0010] Oxidative damage causes significant deterioration of red blood cells (RBCs) and related downstream events in conventionally stored blood; therefore, methods are needed to reduce the degree of oxidative stress and thereby mitigate RBC deterioration. Several approaches have been developed to minimize deterioration and improve transfusion outcomes. The approaches include additive solutions (e.g., U.S. Patent No. 4,769,318 for Hamasaki et al., U.S. Patent No. 4,880,786 for Sasakawa et al., and U.S. Patent No. 6,447,987 for Hess et al.), and cryopreservation (U.S. Patent No. 6,413,713 for Serebrennikov, Chaplin et al., “Blood Cells for Transfusion,” Blood, 59:1118-20 (1982), and Valeri et al., “The survival, function, and hemolysis of human RBCs stored at 4 degrees C in additive solution (AS-1, AS-3, or AS-5) for 42 days and then biochemically modified, frozen, thawed, washed, and stored at 4 degrees C in sodium chloride and glucose solution for 24 Examples include hours, see Transfusion, 40:1341-5 (2000) (these are all incorporated herein by reference).

[0011] One approach that has proven successful in improving blood quality and expanding its practical applications is through storage under oxygen-depleted and anaerobic conditions. Benefits of preserving blood under oxygen-depleted conditions include improved levels of ATP and 2,3-DPG, as well as reduced hemolysis. U.S. Patents 5,624,794, 6,162,396, and 5,476,764 (all incorporated herein by reference) pertain to the preservation of red blood cells under oxygen-depleted conditions. U.S. Patent 5,789,151 (all incorporated herein by reference) pertains to blood preservation additive solutions. U.S. Patent No. 6,162,396 (396 patent) to Bitensky et al. (the entire patent is incorporated herein by reference) discloses an anaerobic storage bag for blood storage, comprising an oxygen-impermeable outer layer and an oxygen-permeable erythrocyte (RBC)-compatible inner layer, with an oxygen scrubber positioned between the inner and outer layers.

[0012] Furthermore, storing blood under oxygen-depleted conditions may result in a reduction in microparticle levels, reduced loss of deformability, reduced oxidation of lipids and proteins, and increased post-transfusion survival compared to blood stored under conventional conditions. See Yoshida et al., “The effects of additive solution pH and metabolic rejuvenation on anaerobic storage of red cells,” Transfusion 48:2096-2105 (2008) and Yoshida, T., et al. “Reduction of microparticle generation during anaerobic storage of red blood cells. Transfusion”, 52, 83A (2012) (these works are incorporated herein by reference in their entirety). Anaerobic storage of red blood cells also results in increased 24-hour in vivo recovery rates after autologous transfusion, increased 2,3-DPG and ATP levels, reduced hemolysis, and beneficial regeneration in metabolic pathways. See Reisz et al., “Oxidative modifications of glyceraldehyde 3-phosphate dehydrogenase regulate metabolic reprogramming of stored red blood cells,” Blood, 128:e32-42 (2016); and Yoshida et al., “Extended storage of red blood cells under anaerobic conditions,” Vox sanguinis 92:22-31 (2007) (these are incorporated herein by reference in their entirety).

[0013] In this disclosure, the inventors demonstrate that oxygen-reduced (OR) or oxygen and carbon dioxide-reduced (OCR) blood from rats results in improved ATP and 2,3-DPG levels during storage compared to conventionally stored blood, as demonstrated in the past using human blood. Therefore, OR or OCR rat RBCs are expected to similarly reduce microparticles, improve deformability, reduce lipid and protein oxidation, and increase post-transfusion survival rates.

[0014] In this specification, the inventors demonstrate for the first time that OR and OCR blood provide remarkable improvements in clinical outcomes when transfused in rats to treat hemorrhagic trauma. Using a rat hemorrhagic shock resuscitation model, the inventors show that OR or OCR RBCs provide reduced organ damage compared to conventionally stored blood. In addition, OR or OCR RBCs provide reversal of shock using a smaller volume of pRBCs. Finally, OR or OCR RBCs stabilized hemodynamics more rapidly than conventionally stored pRBCs of the same storage period when transfused to treat hemorrhagic shock.

[0015] OR and OCR RBCs offer an improved method for treating trauma resulting in blood loss, reducing mortality and pathological conditions compared to conventional stored blood. OR and OCR RBCs provide a reduction in organ failure, including a reduction in marker levels for lung and liver injury. Furthermore, OR and OCR RBCs provide a reduction in the amount of blood required for hemodynamic function recovery and stabilization. Therefore, OR and OCR RBCs can provide a reduction in the amount of RBCs required for transfusion therapy when treating hemorrhagic trauma. The quality improvements of OR and OCR also provide unexpected reductions in trauma-related organ injury, pathological conditions, and mortality, in addition to the previously demonstrated improvements in oxygen delivery capacity of stored RBCs. [Overview of the project]

[0016] This disclosure provides, and includes, a method for treating low mean arterial pressure in a subject requiring treatment for low mean arterial pressure, comprising supplying oxygen-depleted stored blood having an oxygen saturation of 20% or less before and during storage, wherein, after supplying the oxygen-depleted blood to the subject requiring treatment, the mean arterial pressure in the subject requiring treatment increases, and the low mean arterial pressure is due to hemorrhagic trauma.

[0017] This disclosure provides, and includes, a method for reducing the amount of blood required for transfusion in trauma patients requiring transfusion, comprising supplying oxygen-reduced blood having an oxygen saturation of 20% or less before and during storage.

[0018] This disclosure provides, and includes, a method for reducing hemorrhagic shock in a trauma patient requiring reduction of hemorrhagic shock, comprising supplying deoxygenated blood having an oxygen saturation of 20% or less before and during storage, wherein the trauma patient has a lactate level between 0.5 and 2.5 mmol / L per liter before such supply, and the hemorrhagic shock is reversed.

[0019] This disclosure provides, and includes, a method for reducing liver injury in trauma patients requiring blood transfusion therapy, comprising supplying oxygen-depleted blood having an oxygen saturation of 20% or less before and during storage.

[0020] This disclosure provides, and includes, a method for reducing renal failure in patients with hemorrhagic trauma requiring blood transfusion therapy, comprising supplying oxygen-depleted blood having an oxygen saturation of 20% or less before and during storage.

[0021] This disclosure provides, and includes, a method for reducing lung injury in patients with hemorrhagic trauma requiring blood transfusion therapy, comprising supplying oxygen-depleted blood having an oxygen saturation of 20% or less before and during storage.

[0022] This disclosure is provided with reference to the attached drawings.

Brief Description of the Drawings

[0023] [Figure 1] A graph presenting the results of exemplary embodiments according to the present disclosure, comparing the ATP levels in conventional stored RBCs (untreated; control), sham controls (SC), oxygen-reduced RBCs (N2; OR), and oxygen- and carbon dioxide-reduced RBCs (CO2; OCR). [Figure 2] A graph presenting the results of exemplary embodiments according to the present disclosure, comparing the 2,3-DPG levels in conventional stored RBCs (untreated; control), sham controls (SC), oxygen-reduced RBCs (N2; OR), and oxygen- and carbon dioxide-reduced RBCs (CO2; OCR). [Figure 3] A graph presenting the results of exemplary embodiments according to the present disclosure, showing a comparison of the percentage recovery of control, sham, OR RBCs, and OCR RBCs transfused into animals. [Figure 4A] Figures 4A and 4B are graphs presenting the results of exemplary embodiments according to the present disclosure, showing a comparison of the hematocrit percentages in animals resuscitated using control, OR-RBC, and OCR-RBC stored for 1 week (Figure 4A) or 3 weeks (Figure 4B). BL (baseline) identifies animals not in a shock state. Shock identifies animals under hemorrhagic shock. Early R identifies a 10-minute resuscitation period. Late R identifies a 60-minute resuscitation period. [Figure 4B] Same as above. [Figure 5A] Figures 5A and 5B are graphs presenting the results of exemplary embodiments according to the present disclosure, showing a comparison of the mean arterial pressure (MAP) in animals resuscitated using control, OR-RBC, and OCR-RBC stored for 1 week (Figure 5A) or 3 weeks (Figure 5B). BL (baseline) identifies animals not in a shock state. Shock identifies animals under hemorrhagic shock. Early R identifies a 10-minute resuscitation period. Late R identifies a 60-minute resuscitation period. [Figure 5B] Same as above. [Figure 6A] Figures 6A and 6B are graphs presenting the results of exemplary embodiments according to the present disclosure, showing a comparison of the percentage of blood volume supplied to animals during resuscitation after 10, 20, 30, 45, and 60 minutes. Controls, OR-RBCs, and OCR-RBCs stored for 1 week (Figure 6A) or 3 weeks (Figure 6B) are compared. [Figure 6B] The same as above. [Figure 7A] Figures 7A and 7B are graphs presenting the results of exemplary embodiments according to the present disclosure, showing a comparison of the amount of lactate in animals resuscitated using controls, OR-RBCs, and OCR-RBCs stored for 1 week (Figure 7A) or 3 weeks (Figure 7B). BL (baseline) identifies animals not in a shock state. Shock identifies animals under hemorrhagic shock. Early R identifies a 10-minute resuscitation period. Late R identifies a 60-minute resuscitation period. [Figure 7B] The same as above. [Figure 8A] Figures 8A and 8B are graphs presenting the results of exemplary embodiments according to the present disclosure, showing a comparison of the amount of glucose in animals resuscitated using controls, OR-RBCs, and OCR-RBCs stored for 1 week (Figure 8A) or 3 weeks (Figure 8B). BL (baseline) identifies animals not in a shock state. Shock identifies animals under hemorrhagic shock. Early R identifies a 10-minute resuscitation period. Late R identifies a 60-minute resuscitation period. [Figure 8B] The same as above. [Figure 9A] Figures 9A and 9B are graphs presenting the results of exemplary embodiments according to the present disclosure, showing a comparison of the amount of AST in animals resuscitated using controls, OR-RBCs, and OCR-RBCs stored for 1 week (Figure 9A) or 3 weeks (Figure 9B). [Figure 9B] The same as above. [Figure 10A]Figures 10A and 10B are graphs illustrating the results of exemplary embodiments of the present disclosure, showing a comparison of ALT levels in animals preserved for one week (Figure 10A) or three weeks (Figure 10B), OR-RBC, and animals resuscitated using OCR-RBC. [Figure 10B] Same as above. [Figure 11A] Figures 11A and 11B are graphs illustrating the results of exemplary embodiments of the present disclosure, showing a comparison of serum creatinine levels in control, OR-RBC, and OCR-RBC animals preserved for one week (Figure 11A) or three weeks (Figure 11B). [Figure 11B] Same as above. [Figure 12A] Figures 12A and 12B are graphs illustrating the results of exemplary embodiments of the present disclosure, showing a comparison of blood urea nitrogen (BUN) levels in control, OR-RBC, and animals resuscitated using OCR-RBC after being stored for one week (Figure 12A) or three weeks (Figure 12B). [Figure 12B] Same as above. [Figure 13A] Figures 13A and 13B are graphs illustrating the results of exemplary embodiments of the present disclosure, showing a comparison of CXCL1 levels in the livers of control, OR-RBC, and animals resuscitated using OCR-RBC after being stored for one week (Figure 13A) or three weeks (Figure 13B). [Figure 13B] Same as above. [Figure 14A] Figures 14A and 14B are graphs illustrating the results of exemplary embodiments of the present disclosure, showing a comparison of CXCL1 levels in the spleens of control, OR-RBC, and animals resuscitated using OCR-RBC after being stored for one week (Figure 14A) or three weeks (Figure 14B). [Figure 14B] Same as above. [Figure 15A] Figures 15A and 15B are graphs illustrating the results of exemplary embodiments of the present disclosure, showing a comparison of the amount of CXCL1 in the lungs of control, OR-RBC, and animals resuscitated using OCR-RBC after being stored for one week (Figure 15A) or three weeks (Figure 15B). [Figure 15B] Same as above. [Figure 16A] Figures 16A and 16B are graphs illustrating the results of exemplary embodiments of the present disclosure, showing a comparison of urinary neutrophil gelatinase-associated lipocalin (u-NGAL) levels in control, OR-RBC, and animals resuscitated using OCR-RBC after being stored for one week (Figure 16A) or three weeks (Figure 16B). [Figure 16B] Same as above. [Figure 17A] Figures 17A and 17B are graphs illustrating the results of exemplary embodiments of the present disclosure, showing a comparison of the percentage of CD45+ neutrophils in animals resuscitated using control, OR-RBC, and OCR-RBC, stored for one week (Figure 17A) or three weeks (Figure 17B). [Figure 17B] Same as above. [Figure 18A] Figures 18A and 18B are graphs illustrating the results of exemplary embodiments of the present disclosure, showing a comparison of IL-6 levels in control, OR-RBC, and OCR-RBC animals preserved for one week (Figure 18A) or three weeks (Figure 18B). [Figure 18B] Same as above. [Modes for carrying out the invention]

[0024] The embodiments described herein illustrate some of the embodiments of the disclosure, but should not be construed as limiting the scope of the disclosure in any way.

[0025] The present disclosure provides, and includes, a method for supplying a patient with hemorrhagic trauma with oxygen-reduced storage blood having an oxygen saturation of 20% or less before and during storage. The method also provides supplying a patient with hemorrhagic trauma with oxygen-reduced storage blood having an oxygen saturation of 15-20% before and during storage. The method also provides supplying a patient with hemorrhagic trauma with oxygen-reduced storage blood having an oxygen saturation of 10-15% before and during storage. The method also provides supplying a patient with hemorrhagic trauma with oxygen-reduced storage blood having an oxygen saturation of 5-10% before and during storage. The method also provides supplying a patient with hemorrhagic trauma with oxygen-reduced storage blood having an oxygen saturation of 3-5% before and during storage.

[0026] The method also provides supplying oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage for transfusion to persons with hemorrhagic shock. The method also provides supplying oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage for transfusion to persons with hemorrhagic trauma. The method also includes transfusing oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage to patients whose risk of trauma is increased by surgery. A method for supplying oxygen-reduced stored blood having an initial oxygen saturation of 20% or less includes supplying oxygen-reduced stored blood having an initial oxygen saturation of 10% or less. A method for supplying oxygen-reduced stored blood having an initial oxygen saturation of 20% or less further includes supplying oxygen-reduced stored blood having an initial oxygen saturation of 5% or less. A method for supplying oxygen-reduced stored blood having an initial oxygen saturation of 20% or less further includes supplying oxygen-reduced stored blood having an initial oxygen saturation of 3% or less.

[0027] The method of this disclosure provides, and includes, supplying oxygen-reduced preserved blood for the treatment of trauma, wherein the oxygen-reduced preserved blood has an oxygen saturation of 20% or less before and during a preservation period of at least one week, at least two weeks, at least three weeks, at least four weeks, at least five weeks, or at least six weeks. The method also provides supplying oxygen-reduced preserved blood for the treatment of trauma, wherein the oxygen-reduced preserved blood has an oxygen saturation of 15% or less after a preservation period of at least one week, at least two weeks, at least three weeks, at least four weeks, at least five weeks, or at least six weeks. The method also provides supplying oxygen-reduced preserved blood for the treatment of trauma, wherein the oxygen-reduced preserved blood has an oxygen saturation of 10% or less after a preservation period of at least one week, at least two weeks, at least three weeks, at least four weeks, at least five weeks, or at least six weeks. The method further provides supplying oxygen-reduced preserved blood for the treatment of trauma, wherein the oxygen-reduced preserved blood has an oxygen saturation of 5% or less after a storage period of at least 1 week, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, or at least 6 weeks. The method further provides supplying oxygen-reduced preserved blood for the treatment of trauma, wherein the oxygen-reduced preserved blood has an oxygen saturation of 3% or less after a storage period of at least 1 week, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, or at least 6 weeks. The method also provides supplying oxygen-reduced preserved blood for the treatment of trauma, wherein the oxygen-reduced preserved blood has an oxygen saturation of between 3% and 5% after a storage period of at least 1 week, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, or at least 6 weeks. The method also provides supplying oxygen-reduced preserved blood for the treatment of trauma, wherein the oxygen-reduced preserved blood has an oxygen saturation of between 5 and 10% after a preservation period of at least one week, at least two weeks, at least three weeks, at least four weeks, at least five weeks, or at least six weeks.The method also provides supplying oxygen-reduced preserved blood for the treatment of trauma, wherein the oxygen-reduced preserved blood has an oxygen saturation of 10-15% after a storage period of at least 1 week, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, or at least 6 weeks. The method also provides supplying oxygen-reduced preserved blood for the treatment of trauma, wherein the oxygen-reduced preserved blood has an oxygen saturation of 15-20% after a storage period of at least 1 week, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, or at least 6 weeks.

[0028] The method of this disclosure provides, and includes, supplying a trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the trauma patient has suffered a head injury, a penetrating wound, a blunt instrument injury, an injury from a fall, or an injury from a car accident. In another embodiment, the trauma patient is a hemorrhagic trauma patient. In yet another embodiment, the trauma patient is bleeding due to surgery, a penetrating wound, a blunt instrument injury, an injury from a fall, or an injury from a car accident.

[0029] In one embodiment of this disclosure, a trauma patient or a hemorrhagic trauma patient is subject to requiring OR and OCR-stored blood. In one embodiment of this disclosure, a trauma patient is a hemorrhagic trauma patient requiring one or more units of blood as transfusion therapy. In one embodiment of this disclosure, a trauma patient is a hemorrhagic trauma patient requiring two or more units of blood as transfusion therapy. In one embodiment of this disclosure, a trauma patient is a hemorrhagic trauma patient requiring three or more units of blood as transfusion therapy.

[0030] In one embodiment of this disclosure, the trauma patient is a patient in a state of hemorrhagic shock. In one embodiment, the trauma patient is in a state of hemorrhagic shock due to head trauma, a penetrating wound, blunt force trauma, injury from a fall, or injury from a car accident. In one embodiment of this disclosure, the hemorrhagic trauma patient is a patient with Class I bleeding. In another embodiment, the hemorrhagic trauma patient is a patient with Class II bleeding. In another embodiment, the hemorrhagic trauma patient is a patient with Class III bleeding. In another embodiment, the hemorrhagic trauma patient is a patient with Class IV bleeding. In one embodiment of this disclosure, the hemorrhagic trauma patient loses up to 15% of their blood volume. In another embodiment, the hemorrhagic trauma patient loses between 15% and 30% of their blood volume. In another embodiment, the hemorrhagic trauma patient loses between 30% and 40% of their blood volume. In another embodiment, the hemorrhagic trauma patient loses more than 40% of their blood volume.

[0031] This disclosure provides, and includes, that patients requiring transfusion therapy with OR or OCR RBCs exhibit one or more signs selected from the group consisting of decreased mean arterial pressure, decreased hematocrit, increased lactate, increased glucose, increased aspartate aminotransferase (AST), increased alanine aminotransferase (ALT), increased urinary neutrophil gelatinase-associated lipocalin (u-NGAL), increased serum creatinine, and increased blood urea nitrogen. In one aspect of this disclosure, patients requiring transfusion therapy with OR or OCR RBCs are hemorrhagic trauma patients with decreased mean arterial pressure. This disclosure provides, and includes, that patients requiring transfusion therapy with OR or OCR RBCs exhibit increased aspartate aminotransferase (AST) and increased alanine aminotransferase (ALT). This disclosure provides, and includes, that patients requiring transfusion therapy with OR or OCR RBCs exhibit decreased mean arterial pressure and increased lactate. This disclosure provides, and includes, that patients requiring transfusion therapy with OR or OCR RBCs exhibit increased aspartate aminotransferase (AST), increased alanine aminotransferase (ALT), and increased blood urea nitrogen. This disclosure provides, and includes, that patients requiring transfusion therapy with OR or OCR RBCs exhibit increased aspartate aminotransferase (AST), increased alanine aminotransferase (ALT), increased serum creatinine, and increased blood urea nitrogen. This disclosure provides, and includes, that patients requiring transfusion therapy with OR or OCR RBCs exhibit increased lactate and increased glucose. This disclosure provides, and includes, that patients requiring transfusion therapy with OR or OCR RBCs exhibit increased urinary neutrophil gelatinase-associated lipocalin (u-NGAL), increased serum creatinine, and increased blood urea nitrogen.

[0032] In another embodiment, patients requiring transfusion therapy with OR or OCR RBCs are hemorrhagic trauma patients with reduced hematocrit. In another embodiment, patients requiring transfusion therapy with OR or OCR RBCs are hemorrhagic trauma patients with elevated lactate. In yet another embodiment, patients requiring transfusion therapy with OR or OCR RBCs are hemorrhagic trauma patients with elevated glucose. In a further embodiment, hemorrhagic trauma patients with elevated aspartate aminotransferase (AST). In another embodiment, patients requiring transfusion therapy with OR or OCR RBCs are hemorrhagic trauma patients with elevated alanine aminotransferase (ALT). In another embodiment, patients requiring transfusion therapy with OR or OCR RBCs are hemorrhagic trauma patients with elevated urinary neutrophil gelatinase-associated lipocalin (u-NGAL). In another embodiment, patients requiring transfusion therapy with OR or OCR RBCs are hemorrhagic trauma patients with elevated serum creatinine. In another embodiment, patients requiring transfusion therapy with OR or OCR RBCs are hemorrhagic trauma patients with elevated blood urea nitrogen.

[0033] In one embodiment of this disclosure, OR and OCR-stored blood for use in transfusion therapy for trauma patients requiring transfusion therapy has an initial oxygen saturation of 20% or less. In another embodiment, OR and OCR-stored blood has an initial oxygen saturation of 10% or less. In another embodiment, OR and OCR-stored blood has an initial oxygen saturation of 5% or less. In yet another embodiment, OR and OCR-stored blood has an initial oxygen saturation of 3% or less.

[0034] In one embodiment of this disclosure, OCR-stored blood for use in transfusion therapy for trauma patients requiring transfusion therapy has an initial pCO2 (at 37°C) between 10 and 40 mmHg. In another embodiment, OCR-stored blood has an initial pCO2 between 10 and 30 mmHg. In yet another embodiment, OCR-stored blood has an initial pCO2 between 10 and 20 mmHg. In yet another embodiment, OCR-stored blood has an initial pCO2 between 10 and 15 mmHg. In yet another embodiment, OCR-stored blood has an initial pCO2 of less than 10 mmHg.

[0035] In one aspect of this disclosure, OR and OCR-stored blood for use in transfusion therapy for trauma patients requiring transfusion therapy has an initial oxygen saturation of 20% or less and is stored for less than 2 days. In one aspect, OR and OCR-stored blood has an initial oxygen saturation of 20% or less and is stored for less than 7 days. In another aspect, OR and OCR-stored blood has an initial oxygen saturation of 20% or less and is stored for less than 14 days. In another aspect, oxygen-reduced stored blood has an initial oxygen saturation of 20% or less and is stored for less than 21 days. In another aspect, oxygen-reduced stored blood for use in transfusion therapy for trauma patients requiring transfusion therapy has an initial oxygen saturation of 20% or less and is stored for less than 28 days. In another aspect, oxygen-reduced stored blood has an initial oxygen saturation of 20% or less and is stored for less than 35 days. In another aspect, oxygen-reduced stored blood has an initial oxygen saturation of 20% or less and is stored for less than 42 days. In another aspect, oxygen-reduced stored blood has an initial oxygen saturation of 20% or less and is stored for less than 45 days. In one embodiment of this disclosure, OR and OCR-stored blood have an oxygen saturation of 20% or less during storage.

[0036] Suitable blood for use in the method for use in transfusion therapy for trauma patients requiring transfusion therapy in this disclosure includes oxygen-depleted preserved blood containing an anticoagulant. In one embodiment of this disclosure, oxygen-depleted red blood cells are preserved for up to three weeks to produce oxygen-depleted preserved blood. In another embodiment, oxygen-depleted preserved blood usually further includes an additive solution. Suitable additive solutions in this disclosure include AS-1, AS-3 (Nutricel®), AS-5, SAGM, PAGG-SM, PAGG-GM, MAP, AS-7, ESOL-5, EAS61, OFAS1, OFAS3, and combinations thereof. In one embodiment, the additive solution is added during the separation of the components. In one embodiment, the additive solution is AS-1. In another embodiment, the additive solution is AS-3. In yet another embodiment, the additive solution is SAGM.

[0037] The methods of this disclosure provide, and include, increasing the mean arterial pressure (MAP) in trauma patients requiring transfusion therapy, comprising supplying the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the mean arterial pressure increases by between 20% and 60%. In another embodiment, the mean arterial pressure increases by between 30% and 60%. In yet another embodiment, the mean arterial pressure of trauma patients receiving transfusion therapy with OR or OCR blood increases by between 30% and 50%. In yet another embodiment, the mean arterial pressure increases by between 30% and 60%. In a further embodiment, the mean arterial pressure of trauma patients receiving transfusion therapy with OR or OCR blood increases by between 30% and 40%. In one embodiment, the mean arterial pressure increases by at least 10, 20, 30, 40, 50, 60, 70, 80, or 90% compared to the mean arterial pressure of a patient transfused with conventional stored blood.

[0038] In one aspect of this disclosure, the mean arterial pressure increases by at least 1.5 times. In another aspect, the mean arterial pressure of trauma patients receiving transfusion therapy with OR or OCR blood increases by at least 2 times. In a further aspect, the mean arterial pressure increases between 1 and 2 times. In one aspect of this disclosure, the mean arterial pressure of trauma patients receiving transfusion therapy with OR or OCR blood increases by at least 10 mmHg, at least 20 mmHg, at least 30 mmHg, at least 40 mmHg, at least 50 mmHg, or at least 60 mmHg. In another aspect, the mean arterial pressure of trauma patients receiving transfusion therapy with OR or OCR blood increases between 20 and 50 mmHg. In a further aspect, the mean arterial pressure increases between 30 and 50 mmHg.

[0039] The present disclosure provides, and includes, a method for increasing the mean arterial pressure in a trauma patient requiring transfusion therapy to between 70 and 110 mmHg, comprising supplying the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In another embodiment, the mean arterial pressure of a trauma patient receiving transfusion therapy with OR or OCR blood is increased to at least 70 mmHg. In another embodiment, the mean arterial pressure of a trauma patient receiving transfusion therapy with OR or OCR blood is increased to at least 80 mmHg. In yet another embodiment, the mean arterial pressure is increased to at least 90 mmHg. In a further embodiment, the mean arterial pressure is increased to at least 100 mmHg. In one embodiment of the present disclosure, the mean arterial pressure in a subject requiring it is maintained between 70 and 110 mmHg for at least 1 hour after transfusion. In another embodiment, the mean arterial pressure is maintained between 70 and 110 mmHg for at least 2 hours after transfusion. In another embodiment, mean arterial pressure is maintained between 70 and 105 mmHg for at least 3 hours after transfusion. In another embodiment, mean arterial pressure is maintained between 70 and 110 mmHg for at least 4 hours after transfusion. In yet another embodiment, mean arterial pressure is maintained between 70 and 110 mmHg for at least 5 hours after transfusion.

[0040] The present disclosure provides, and includes, a method for increasing the mean arterial pressure in trauma patients requiring transfusion therapy at a faster rate than in patients transfused with conventionally stored blood, comprising supplying the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the mean arterial pressure of patients transfused with OR or OCR blood recovers to normal physiological parameters in half the time compared to conventionally stored blood.

[0041] The method of this disclosure provides, and includes, a reduction in the amount of stored blood required for transfusion in trauma patients requiring transfusion therapy, comprising supplying the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the amount of OR stored blood required for transfusion is 10 to 90% less than the amount of conventional stored blood required. In another embodiment, the amount of OR stored blood required for transfusion is 10 to 30% less than the amount of conventional stored blood required. In another embodiment, the amount of OR stored blood required for transfusion is 20 to 50% less than the amount of conventional stored blood required. In another embodiment, the amount of OR stored blood required for transfusion is 20 to 80% less than the amount of conventional stored blood required. In yet another embodiment, the amount of OR stored blood required for transfusion is 30 to 80% less than the amount of conventional stored blood required. In yet another embodiment, the amount of OR stored blood required for transfusion is 40 to 85% less than the amount of conventional stored blood required. In a further embodiment, the amount of OR-stored blood required for transfusion is 50-90% less than the amount of conventionally stored blood required.

[0042] The method of this disclosure provides, and includes, reducing the amount of stored blood required for transfusion in trauma patients requiring transfusion therapy by at least 10%, comprising supplying the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the amount of OR stored blood required for transfusion is at least 20% less than the amount of conventional stored blood required. In another embodiment, the amount of OR stored blood required for transfusion is at least 30% less than the amount of conventional stored blood required. In another embodiment, the amount of OR stored blood required for transfusion is at least 40% less than the amount of conventional stored blood required. In another embodiment, the amount of OR stored blood required for transfusion is at least 50% less than the amount of conventional stored blood required. In yet another embodiment, the amount of OR stored blood required for transfusion is at least 60% less than the amount of conventional stored blood required. In yet another embodiment, the amount of OR stored blood required for transfusion is at least 70% less than the amount of conventional stored blood required. In a further embodiment, the amount of OR-stored blood required for transfusion is about 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-95% less than the amount of conventionally stored blood required. In another embodiment, the amount of OR-stored blood required for transfusion therapy in trauma patients requiring transfusion therapy is about 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-95% less than the amount of conventionally stored blood required.

[0043] Lactate clearance is a biomarker for resuscitation from hemorrhagic shock. See: Hashmi et al., “Predictors of mortality in geriatric trauma patients: a systematic review and meta-analysis,” The Journal of Trauma and Acute Care Surgery, 76:894-901 (2014); Regnier et al., “Prognostic significance of blood lactate and lactate clearance in trauma patients,” Anesthesiology, 117:1276-88 (2012); and Zhang et al., “Lactate clearance is a useful biomarker for the prediction of all-cause mortality in critically ill patients: a systematic review and meta-analysis,” Critical Care Medicine, 42:2118-25 (2014) ("Zhang 2014") (these are incorporated herein by reference in their entirety). Clinical values ​​of lactate clearance are useful in predicting outcomes in patients with septic shock and critical conditions without apparent circulatory shock. Elevated lactate levels are an indicator of adverse clinical outcomes, and rapid lactate clearance has been widely associated with improved outcomes in diverse ICU or ED patient populations. See Zhang 2014. The reduction in lactate levels in animals resuscitated with OR-RBCs compared to conventional RBCs supports the idea that resuscitation with OR RBCs can significantly improve patient clinical outcomes. See Figures 7A and 7B.

[0044] The methods of this disclosure provide, and include, reducing lactate levels in trauma patients requiring transfusion therapy, comprising supplying the trauma patient with oxygen-reduced (OR) stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the lactate level is reduced to between 10% and 90%. In one embodiment, transfusion using OR stored blood reduces the lactate level in trauma patients requiring transfusion therapy to between 10% and 50%. In another embodiment, transfusion using OR stored blood reduces the lactate level in trauma patients requiring transfusion therapy to between 20% and 40%. In another embodiment, transfusion using OR stored blood reduces the lactate level in trauma patients requiring transfusion therapy to between 50% and 90%. In yet another embodiment, transfusion using OR stored blood reduces the lactate level in trauma patients requiring transfusion therapy to between 60% and 90%. In another embodiment, transfusion using OR-stored blood reduces lactate levels in trauma patients requiring transfusion therapy by 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, or 80-90%. In another embodiment, transfusion using OR-stored blood reduces lactate levels in trauma patients requiring transfusion therapy by at least 10%. In another embodiment, transfusion using OR-stored blood reduces lactate levels in trauma patients requiring transfusion therapy by at least 20%. In a further embodiment, transfusion using OR-stored blood reduces lactate levels in trauma patients requiring transfusion therapy by at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90%.

[0045] The present disclosure provides, and includes, a method for reducing elevated lactate levels in trauma patients requiring transfusion therapy to between approximately 0.5 and approximately 2.5 mmol / L, comprising supplying the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the lactate level in a trauma patient requiring transfusion therapy is reduced to between approximately 0.9 and approximately 2 mmol / L. In another embodiment, the lactate level in a trauma patient requiring transfusion therapy is reduced to between approximately 0.9 and approximately 1.7 mmol / L. In yet another embodiment, the lactate level in a patient requiring transfusion therapy is reduced to between approximately 1.4 and approximately 2.4 mmol / L. In yet another embodiment, the lactate level in a trauma patient requiring transfusion therapy is reduced to between approximately 1.7 and approximately 2.5 mmol / L. In yet another embodiment, the lactate level in a trauma patient requiring transfusion therapy is reduced to less than approximately 2.5 mmol / L. In a further embodiment, the lactate level in trauma patients requiring blood transfusion is reduced to less than approximately 2.0 mmol / L. In another embodiment, the lactate level in trauma patients requiring blood transfusion is reduced to less than approximately 1.5 mmol / L. In yet another embodiment, the lactate level in trauma patients requiring blood transfusion is reduced to less than approximately 1.0 mmol / L. In yet another embodiment, the lactate level in trauma patients requiring blood transfusion is reduced to between approximately 0.5 and approximately 1.0 mmol / L.

[0046] The present disclosure provides, and includes, a method for reducing elevated lactate levels in trauma patients requiring transfusion therapy to between 0.5 and 2.5 mmol / L, comprising supplying the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the lactate level in a patient requiring transfusion therapy is reduced to between 0.9 and 2 mmol / L. In another embodiment, the lactate level in a trauma patient requiring transfusion therapy is reduced to between 0.9 and 1.7 mmol / L. In yet another embodiment, the lactate level in a trauma patient requiring transfusion therapy is reduced to between 1.4 and 2.4 mmol / L. In yet another embodiment, the lactate level in a trauma patient requiring transfusion therapy is reduced to between 1.7 and 2.5 mmol / L. In yet another embodiment, the lactate level in a trauma patient requiring transfusion therapy is reduced to between 0.5 and 1 mmol / L.

[0047] The present disclosure provides, and includes, a method for reducing elevated lactate levels in hemorrhagic trauma patients requiring transfusion therapy to less than 4 mmol / L, comprising supplying the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the lactate level in a trauma patient requiring transfusion therapy is reduced to less than 3 mmol / L. In another embodiment, the lactate level in a trauma patient requiring transfusion therapy is reduced to less than 2.5 mmol / L. In another embodiment, the lactate level in the patient is reduced to less than 2.3 mmol / L. In another embodiment, the lactate level in a trauma patient requiring transfusion therapy is reduced to less than 2 mmol / L. In another embodiment, the lactate level in a trauma patient requiring transfusion therapy is reduced to less than 2 mmol / L. In another embodiment, the lactate level in a trauma patient requiring transfusion therapy is reduced to less than 1.5 mmol / L. In another embodiment, lactate levels in trauma patients requiring blood transfusions are reduced to less than 1 mmol / L.

[0048] Blood glucose levels are also known to be a predictor of outcomes in several disease patterns, particularly in trauma patients. Trauma patients are more likely to have a poorer outcome due to hyperglycemia than patients with other severe conditions. See Kreutziger et al., “Admission blood glucose predicted hemorrhagic shock in multiple trauma patients,” Injury, 46:15-20 (2015) (the entire work is incorporated herein by reference). In studies evaluating the relationship between early hyperglycemia and trauma patients, early hyperglycemia was examined at three possible cutoffs: glucose > 110 mg / dL, glucose > 150 mg / dL, and glucose > 200 mg / dL. See Laird et al., “Relationship of early hyperglycemia to mortality in trauma patients,” J Trauma, 56:1058-62 (2004) (the entire work is incorporated herein by reference). Glucose levels >200 mg / dL were associated with significantly higher infection and mortality rates in trauma patients, regardless of injury characteristics. This was not the case at the 110 mg / dL or 150 mg / dL cutoffs. The reduction in glucose levels in animals resuscitated with OR and OR-RBCs compared to those resuscitated with conventional RBCs supports the idea that resuscitation with OR-RBCs can significantly improve patient clinical outcomes. See Figures 8A and 8B.

[0049] The methods of this disclosure provide, and include, reducing glucose in trauma patients requiring transfusion therapy, comprising supplying trauma patients with oxygen-reduced (OR) stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, glucose is reduced by 10 to 90% compared to transfusion of blood stored under conventional conditions. In one embodiment, transfusion using OR stored blood reduces glucose by 10 to 50% compared to transfusion of blood stored under conventional conditions. In another embodiment, transfusion using OR stored blood reduces glucose by 20 to 40% compared to transfusion of blood stored under conventional conditions. In another embodiment, transfusion using OR stored blood reduces glucose by 50 to 90% compared to transfusion of blood stored under conventional conditions. In yet another embodiment, transfusion using OR stored blood reduces glucose by 60 to 90%. In another embodiment, transfusion using OR-stored blood reduces glucose by 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, or 80-90% compared to transfusion of blood stored under conventional conditions. In another embodiment, transfusion using OR-stored blood reduces glucose by at least 10% compared to transfusion of blood stored under conventional conditions. In yet another embodiment, transfusion using OR-stored blood reduces glucose by at least 20%. In a further embodiment, transfusion using OR-stored blood reduces glucose by at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90%.

[0050] The method of this disclosure provides, and includes, reducing glucose levels in trauma patients requiring transfusion therapy to between approximately 70 and approximately 120 mg / dL, comprising supplying the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, glucose in a patient after transfusion therapy with OR or OCR blood is between approximately 70 and approximately 110 mg / dL. In another embodiment, glucose in a patient after transfusion therapy with OR or OCR blood is between approximately 70 and approximately 100 mg / dL. In another embodiment, glucose in a trauma patient after transfusion therapy with OR or OCR blood is between approximately 90 and approximately 120 mg / dL. In another embodiment, glucose in a trauma patient after transfusion therapy with OR or OCR blood is between approximately 90 and approximately 100 mg / dL.

[0051] The method of this disclosure provides, and includes, reducing glucose levels in trauma patients requiring transfusion therapy to between 70 and 120 mg / dL, comprising supplying the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, glucose in a patient after transfusion therapy with OR or OCR blood is between 70 and 110 mg / dL. In another embodiment, glucose in a patient is between 70 and 100 mg / dL. In yet another embodiment, glucose in a patient is between 90 and 120 mg / dL. In yet another embodiment, glucose in a patient after transfusion therapy with OR or OCR blood is between 90 and 100 mg / dL.

[0052] The method of this disclosure provides, and includes, reducing glucose levels in trauma patients requiring transfusion therapy to less than 120 mg / dL, comprising supplying the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In a further embodiment, glucose in a patient after transfusion therapy with OR or OCR blood is less than 110 mmol / L. In yet another embodiment, glucose in a patient after transfusion therapy with OR or OCR blood is less than 100 mg / dL. In yet another embodiment, glucose in a patient after transfusion therapy with OR or OCR blood is less than 200 mg / dL. In yet another embodiment, glucose in a patient after transfusion therapy with OR or OCR blood is less than 90 mg / dL. In yet another embodiment, glucose in a patient after transfusion therapy with OR or OCR blood is less than 80 mg / dL.

[0053] In one embodiment of this disclosure, trauma patients have an increased risk of complications from transfusion therapy based on their pre-existing or underlying condition. In one embodiment, trauma patients have a pre-existing or underlying condition selected from the group consisting of diabetes mellitus, ischemic heart disease, systemic inflammatory syndrome resulting from trauma or infection, multiple organ failure resulting from trauma or infection, smoke inhalation, and chronic obstructive pulmonary disease, e.g., systemic inflammation due to infection, coagulation disorders, and autoimmune diseases. In another embodiment, trauma patients have one or more pre-existing or underlying conditions selected from the group consisting of diabetes mellitus, ischemic heart disease, systemic inflammatory syndrome resulting from trauma or infection, multiple organ failure resulting from trauma or infection, smoke inhalation, and chronic obstructive pulmonary disease, e.g., systemic inflammation due to infection, coagulation disorders, and autoimmune diseases. In yet another embodiment, trauma patients have two or more pre-existing or underlying conditions selected from the group consisting of diabetes mellitus, ischemic heart disease, systemic inflammatory syndrome resulting from trauma or infection, multiple organ failure resulting from trauma or infection, smoke inhalation, and chronic obstructive pulmonary disease, e.g., systemic inflammation due to infection, coagulation disorders, and autoimmune diseases. In another embodiment, the trauma patient has three or more pre-existing or underlying conditions selected from the group consisting of diabetes mellitus, ischemic heart disease, systemic inflammatory syndrome resulting from trauma or infection, multiple organ failure resulting from trauma or infection, smoke inhalation, chronic obstructive pulmonary disease, for example, systemic inflammation due to infection, coagulation disorders, and autoimmune diseases.

[0054] Patients may experience adverse events during hemorrhagic shock, including liver injury or failure, kidney injury or failure, lung injury or failure, or a combination thereof. This disclosure provides, and includes, that patients requiring transfusion therapy with OR or OCR RBCs may exhibit one or more adverse events selected from the group consisting of liver injury or failure, kidney injury or failure, or lung injury or failure. This disclosure provides, and includes, that patients requiring transfusion therapy with OR or OCR RBCs may exhibit two or more adverse events selected from the group consisting of liver injury or failure, kidney injury or failure, or lung injury or failure.

[0055] The methods of this disclosure provide, and include, reducing adverse events in trauma patients, which include supplying trauma patients requiring transfusion therapy with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, adverse events after transfusion using OR or OCR blood are reduced by at least 5%. In another embodiment, adverse events after transfusion using OR or OCR blood are reduced by at least 10%. In another embodiment, adverse events after transfusion using OR or OCR blood are reduced by at least 20%. In another embodiment, adverse events after transfusion using OR or OCR blood are reduced by at least 30%. In another embodiment, adverse events after transfusion using OR or OCR blood are reduced by at least 40%. In another embodiment, adverse events after transfusion using OR or OCR blood are reduced by at least 50%. In another embodiment, adverse events after transfusion using OR or OCR blood are reduced by at least 60%. In another embodiment, adverse events are reduced by at least 70%. In another embodiment, adverse events after transfusion using OR or OCR blood are reduced by at least 80%. In yet another embodiment, adverse events are reduced by at least 90%. In a further embodiment, adverse events after transfusion therapy using OR or OCR blood are reduced by between 1-10%, 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-95%. In one embodiment, the adverse event after transfusion using OR or OCR blood is liver injury or liver damage. In another embodiment, the adverse event is lung injury or lung damage. In yet another embodiment, the adverse event is kidney injury or kidney damage. In a further embodiment, the adverse event is liver injury, lung injury, kidney injury, or a combination thereof.

[0056] Elevated levels of liver enzymes (including, but not limited to, aspartate aminotransferase (AST) and alanine aminotransferase (ALT)) indicate any form of liver damage, shock, or injury. The methods of the present disclosure provide, and include, reducing elevated levels of liver enzymes in a trauma patient, comprising supplying the trauma patient with oxygen-depleted preserved blood having an oxygen saturation of 20% or less before and during preservation.

[0057] The methods of this disclosure provide, and include, reducing AST levels in trauma patients requiring transfusion therapy, comprising supplying the trauma patient with deoxygenated stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the AST level is reduced by at least 5% compared to the AST level of a patient transfused with conventional stored blood. In another embodiment, the AST level is reduced by at least 10% compared to the AST level of a patient transfused with conventional stored blood. In another embodiment, the AST level is reduced by at least 20% compared to the AST level of a patient transfused with conventional stored blood. In another embodiment, the AST level is reduced by at least 30% compared to the AST level of a patient transfused with conventional stored blood. In another embodiment, the AST level is reduced by at least 40%. In another embodiment, the AST level is reduced by at least 50% compared to the AST level of a patient transfused with conventional stored blood. In another embodiment, the AST level is reduced by at least 60%. In another embodiment, AST levels are reduced by at least 70% compared to the AST levels of patients transfused with conventionally stored blood. In yet another embodiment, AST levels are reduced by at least 80%. In a further embodiment, AST levels are reduced by at least 90% compared to the AST levels of patients transfused with conventionally stored blood. In a further embodiment, AST levels are reduced by between 1-10%, 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-95% compared to the AST levels of patients transfused with conventionally stored blood.

[0058] The present disclosure provides, and includes, a method for reducing AST levels in trauma patients requiring transfusion therapy by 1.5 to 10 times, comprising supplying the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the AST level is reduced by 2 to 3 times compared to the AST level of a patient transfused with conventional stored blood. In another embodiment, the AST level is reduced by 3 to 4 times. In another embodiment, the AST level is reduced by 4 to 10 times. In another embodiment, the AST level is reduced by 6 to 9 times compared to the AST level of a patient transfused with conventional stored blood. In a further embodiment, the AST level is reduced by 2 to 5 times. In another embodiment, the AST level is reduced by 10 to 50 times compared to the AST level of a patient transfused with conventional stored blood.

[0059] The method of this disclosure provides, and includes, reducing AST levels by at least 1.5 times in trauma patients requiring blood transfusion therapy, comprising supplying the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the AST level is reduced by at least 2 times compared to the AST level of a patient transfused with conventional stored blood. In another embodiment, the AST level is reduced by at least 3 times compared to the AST level of a patient transfused with conventional stored blood. In another embodiment, the AST level is reduced by at least 4 times compared to the AST level of a patient transfused with conventional stored blood. In another embodiment, the AST level is reduced by at least 5 times compared to the AST level of a patient transfused with conventional stored blood. In a further embodiment, the AST level is reduced by at least 6 times. In another embodiment, the AST level is reduced by at least 7 times compared to the AST level of a patient transfused with conventional stored blood. In another embodiment, the AST level is reduced by at least 8 times. In another embodiment, AST levels are reduced by at least nine times compared to the AST levels of patients who received transfusions using conventionally stored blood. In yet another embodiment, AST levels are reduced by at least ten times compared to the AST levels of patients who received transfusions using conventionally stored blood. In a further embodiment, AST levels are reduced by at least fifty times compared to the AST levels of patients who received transfusions using conventionally stored blood.

[0060] The methods of this disclosure provide, and include, reducing ALT levels in trauma patients requiring transfusion therapy, comprising supplying the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the ALT level is reduced by at least 5% compared to the ALT level of a patient transfused with conventional stored blood. In another embodiment, the ALT level is reduced by at least 10% compared to the ALT level of a patient transfused with conventional stored blood. In another embodiment, the ALT level is reduced by at least 20% compared to the ALT level of a patient transfused with conventional stored blood. In another embodiment, the ALT level is reduced by at least 30% compared to the ALT level of a patient transfused with conventional stored blood. In another embodiment, the ALT level is reduced by at least 40% compared to the AS ALT T level of a patient transfused with conventional stored blood. In another embodiment, the ALT level is reduced by at least 50% compared to the ALT level of a patient transfused with conventional stored blood. In another embodiment, ALT levels are reduced by at least 60% compared to the ALT levels of patients transfused with conventionally stored blood. In yet another embodiment, ALT levels are reduced by at least 70% compared to the ALT levels of patients transfused with conventionally stored blood. In yet another embodiment, ALT levels are reduced by at least 80% compared to the ALT levels of patients transfused with conventionally stored blood. In a further embodiment, ALT levels are reduced by at least 90%. In a further embodiment, ALT levels are reduced by between 1-10%, 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-95% compared to the ALT levels of patients transfused with conventionally stored blood.

[0061] The method of this disclosure provides, and includes, reducing the ALT level in trauma patients requiring transfusion therapy by 1.5 to 10 times, comprising supplying the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the ALT level is reduced by 2 to 3 times compared to the ALT level of a patient transfused with conventional stored blood. In another embodiment, the ALT level is reduced by 3 to 4 times compared to the ALT level of a patient transfused with conventional stored blood. In another embodiment, the ALT level is reduced by 4 to 10 times compared to the ALT level of a patient transfused with conventional stored blood. In yet another embodiment, the ALT level is reduced by 6 to 9 times compared to the ALT level of a patient transfused with conventional stored blood. In a further embodiment, the ALT level is reduced by 2 to 5 times. In another embodiment, ALT levels are reduced by 10 to 50 times compared to the ALT levels of patients who received transfusions using conventionally stored blood.

[0062] The method of this disclosure provides, and includes, reducing the ALT level in a trauma patient requiring transfusion therapy by at least 1.5 times, comprising supplying the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the ALT level is reduced by at least 2 times compared to the ALT level of a patient transfused with conventional stored blood. In another embodiment, the ALT level is reduced by at least 3 times compared to the ALT level of a patient transfused with conventional stored blood. In another embodiment, the ALT level is reduced by at least 4 times compared to the ALT level of a patient transfused with conventional stored blood. In another embodiment, the ALT level is reduced by at least 5 times compared to the ALT level of a patient transfused with conventional stored blood. In a further embodiment, the ALT level is reduced by at least 6 times compared to the ALT level of a patient transfused with conventional stored blood. In another embodiment, the ALT level is reduced by at least 7 times compared to the ALT level of a patient transfused with conventional stored blood. In another embodiment, the ALT level is reduced by at least eight times compared to the ALT level of patients who received transfusions using conventionally stored blood. In yet another embodiment, the ALT level is reduced by at least nine times compared to the ALT level of patients who received transfusions using conventionally stored blood. In yet another embodiment, the ALT level is reduced by at least ten times compared to the ALT level of patients who received transfusions using conventionally stored blood. In a further embodiment, the ALT level is reduced by at least fifty times compared to the ALT level of patients who received transfusions using conventionally stored blood.

[0063] Markers of renal function during and after hemorrhagic trauma include urinary neutrophil gelatinase-associated lipocalin (u-NGAL), serum creatinine, and blood urea nitrogen (BUN). See Treeprasertsuk et al., “Urine neutrophil gelatinase-associated lipocalin: a diagnostic and prognostic marker for acute kidney injury (AKI) in hospitalized cirrhotic patients with AKI-prone conditions,” BMC Gastroenterol 15:140 (2015) (this entire work is incorporated herein by reference). Gene expression analyses reported in more than 150 different studies conducted in AKI models from several species, from rodents to humans, have consistently shown that the NGAL gene is one of the genes most dramatically upregulated in the kidney immediately after an ischemic or nephrotoxic attack. See Ciccia et al., “Pediatric acute kidney injury: prevalence, impact and management challenges,” Int J Nephrol Renovasc Dis, 10:77-84 (2017) (this entire work is incorporated herein by reference). Similarly, serum creatinine levels may vary with age, race, and body size, but elevated creatinine levels indicate kidney injury. Creatinine levels greater than 1.2 in women and greater than 1.4 in men may be an early sign of kidney injury. Elevated blood urea nitrogen (BUN) is seen with kidney disease or renal failure, as well as congestive heart failure, shock, and gastrointestinal bleeding. BUN levels greater than 100 mg / dL indicate severe kidney injury. Decreased BUN levels are also of concern, and may indicate fluid overload, trauma, surgery, opioid use, malnutrition, and anabolic steroid use.See Pagana, “Mosby’s Manual of Diagnostic and Laboratory Tests,” St. Louis Mosby, Inc., (1998); and Gowda, et al., “Markers of renal function tests,” NAm JMed Sci.2(4):170-173(2010) (these are incorporated herein by reference in their entirety). The reductions in u-NGAL (Figures 16A and 16B), serum creatinine (Figures 11A and 11B), and BUN (Figures 12A and 12B) levels in animals resuscitated with OR- and OCR-RBCs compared to conventional RBCs, as provided in this disclosure, demonstrate that resuscitation with OR-RBCs can significantly improve patient clinical outcomes.

[0064] The present disclosure provides a method for reducing urinary neutrophil gelatinase-related lipocalin (u-NGAL) levels in trauma patients requiring transfusion therapy, comprising supplying the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the u-NGAL level is reduced by at least 5% compared to the u-NGAL level of a patient transfused with conventional stored blood. In another embodiment, the u-NGAL level is reduced by at least 10% compared to the u-NGAL level of a patient transfused with conventional stored blood. In another embodiment, the u-NGAL level is reduced by at least 20% compared to the u-NGAL level of a patient transfused with conventional stored blood. In another embodiment, the u-NGAL level is reduced by at least 30%. In another embodiment, the u-NGAL level is reduced by at least 40% compared to the u-NGAL level of a patient transfused with conventional stored blood. In another embodiment, the u-NGAL level is reduced by at least 50% compared to the u-NGAL level of patients who received transfusions using conventionally stored blood. In yet another embodiment, the u-NGAL level is reduced by at least 60% compared to the u-NGAL level of patients who received transfusions using conventionally stored blood. In yet another embodiment, the u-NGAL level is reduced by at least 70% compared to the u-NGAL level of patients who received transfusions using conventionally stored blood. In yet another embodiment, the u-NGAL level is reduced by at least 80% compared to the u-NGAL level of patients who received transfusions using conventionally stored blood. In yet another embodiment, the u-NGAL level is reduced by at least 90% compared to the u-NGAL level of patients who received transfusions using conventionally stored blood. In a further embodiment, u-NGAL levels are reduced by between 1-10%, 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-95% compared to u-NGAL levels in patients transfused with conventionally stored blood.

[0065] The present disclosure provides a method for reducing urinary neutrophil gelatinase-related lipocalin (u-NGAL) levels in trauma patients requiring transfusion therapy, comprising supplying the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage, by a factor of 1.5 to 10. In one embodiment, the u-NGAL level is reduced by a factor of 2 to 3 compared to the u-NGAL level of a patient transfused with conventional stored blood. In another embodiment, the u-NGAL level is reduced by a factor of 3 to 4 compared to the u-NGAL level of a patient transfused with conventional stored blood. In yet another embodiment, the u-NGAL level is reduced by a factor of 4 to 10. In yet another embodiment, the u-NGAL level is reduced by a factor of 6 to 9 compared to the u-NGAL level of a patient transfused with conventional stored blood. In a further embodiment, u-NGAL levels are reduced by 2 to 5 times compared to u-NGAL levels in patients transfused with conventionally stored blood. In another embodiment, u-NGAL levels are reduced by 10 to 50 times compared to u-NGAL levels in patients transfused with conventionally stored blood.

[0066] The present disclosure provides a method for reducing urinary neutrophil gelatinase-associated lipocalin (u-NGAL) levels in trauma patients requiring transfusion therapy, comprising supplying the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage by at least 1.5 times. In one embodiment, the u-NGAL level is reduced by at least 2 times compared to the u-NGAL level of a patient transfused with conventional stored blood. In another embodiment, the u-NGAL level is reduced by at least 3 times compared to the u-NGAL level of a patient transfused with conventional stored blood. In another embodiment, the u-NGAL level is reduced by at least 4 times compared to the u-NGAL level of a patient transfused with conventional stored blood. In another embodiment, the u-NGAL level is reduced by at least 5 times compared to the u-NGAL level of a patient transfused with conventional stored blood. In a further embodiment, the u-NGAL level is reduced by at least six times compared to the u-NGAL level of patients who received transfusions using conventionally stored blood. In another embodiment, the u-NGAL level is reduced by at least seven times compared to the u-NGAL level of patients who received transfusions using conventionally stored blood. In another embodiment, the u-NGAL level is reduced by at least eight times compared to the u-NGAL level of patients who received transfusions using conventionally stored blood. In another embodiment, the u-NGAL level is reduced by at least nine times. In another embodiment, the u-NGAL level is reduced by at least ten times. In a further embodiment, the u-NGAL level is reduced by at least fifty times.

[0067] The present disclosure provides a method for reducing serum creatinine levels in trauma patients requiring blood transfusion therapy, comprising supplying the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the serum creatinine level is reduced by at least 5% compared to the serum creatinine level of a patient transfused with conventional stored blood. In another embodiment, the serum creatinine level is reduced by at least 10% compared to the serum creatinine level of a patient transfused with conventional stored blood. In another embodiment, the serum creatinine level is reduced by at least 20% compared to the serum creatinine level of a patient transfused with conventional stored blood. In another embodiment, the serum creatinine level is reduced by at least 30% compared to the serum creatinine level of a patient transfused with conventional stored blood. In another embodiment, the serum creatinine level is reduced by at least 40% compared to the serum creatinine level of a patient transfused with conventional stored blood. In another embodiment, serum creatinine levels are reduced by at least 50% compared to the serum creatinine levels of patients who received transfusions using conventionally stored blood. In yet another embodiment, serum creatinine levels are reduced by at least 60% compared to the serum creatinine levels of patients who received transfusions using conventionally stored blood. In yet another embodiment, serum creatinine levels are reduced by at least 70% compared to the serum creatinine levels of patients who received transfusions using conventionally stored blood. In yet another embodiment, serum creatinine levels are reduced by at least 80% compared to the serum creatinine levels of patients who received transfusions using conventionally stored blood. In yet another embodiment, serum creatinine levels are reduced by at least 90% compared to the serum creatinine levels of patients who received transfusions using conventionally stored blood. In a further embodiment, serum creatinine levels are reduced by between 1–10%, 10–20%, 20–30%, 30–40%, 40–50%, 50–60%, 60–70%, 70–80%, 80–90%, or 90–95% compared to the serum creatinine levels of patients transfused with conventionally stored blood.

[0068] The present disclosure provides, and includes, a method for reducing serum creatinine levels in trauma patients requiring blood transfusion therapy by 1.5 to 10 times, comprising supplying the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, serum creatinine levels are reduced by 2 to 3 times compared to the serum creatinine levels of patients transfused with conventional stored blood. In another embodiment, serum creatinine levels are reduced by 3 to 4 times compared to the serum creatinine levels of patients transfused with conventional stored blood. In another embodiment, serum creatinine levels are reduced by 4 to 10 times compared to the serum creatinine levels of patients transfused with conventional stored blood. In another embodiment, serum creatinine levels are reduced by 6 to 9 times compared to the serum creatinine levels of patients transfused with conventional stored blood. In a further embodiment, serum creatinine levels are reduced by 2 to 5 times compared to the serum creatinine levels of patients transfused with conventionally stored blood. In another embodiment, serum creatinine levels are reduced by 10 to 50 times compared to the serum creatinine levels of patients transfused with conventionally stored blood.

[0069] The method of this disclosure provides, and includes, reducing serum creatinine levels in trauma patients requiring transfusion therapy by at least 1.5 times, comprising supplying the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, serum creatinine levels are reduced by at least 2 times compared to the serum creatinine levels of patients transfused with conventional stored blood. In another embodiment, serum creatinine levels are reduced by at least 3 times compared to the AST levels of patients transfused with conventional stored blood. In another embodiment, serum creatinine levels are reduced by at least 4 times compared to the serum creatinine levels of patients transfused with conventional stored blood. In yet another embodiment, serum creatinine levels are reduced by at least 5 times. In a further embodiment, serum creatinine levels are reduced by at least 6 times compared to the serum creatinine levels of patients transfused with conventional stored blood. In another embodiment, serum creatinine levels are reduced by at least seven times compared to the serum creatinine levels of patients who received transfusions using conventionally stored blood. In another embodiment, serum creatinine levels are reduced by at least eight times compared to the serum creatinine levels of patients who received transfusions using conventionally stored blood. In another embodiment, serum creatinine levels are reduced by at least nine times compared to the serum creatinine levels of patients who received transfusions using conventionally stored blood. In another embodiment, serum creatinine levels are reduced by at least ten times compared to the serum creatinine levels of patients who received transfusions using conventionally stored blood.

[0070] The present disclosure provides, and includes, a method for reducing serum creatinine levels in trauma patients requiring transfusion therapy to between 0.5 and 1.5 mg / dL, comprising supplying the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the serum creatinine level is reduced to between 0.5 and 1 mg / dL compared to the serum creatinine level of a patient transfused with conventional stored blood. In another embodiment, the serum creatinine level is reduced to between 0.8 and 1 mg / dL compared to the serum creatinine level of a patient transfused with conventional stored blood.

[0071] The present disclosure provides, and includes, a method for reducing serum creatinine levels to less than 1.5 mg / dL in trauma patients requiring blood transfusion therapy, comprising supplying the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the serum creatinine level is reduced to less than 1.4 mg / dL compared to the serum creatinine level of a patient transfused with conventional stored blood. In another embodiment, the serum creatinine level is reduced to less than 1 mg / dL compared to the serum creatinine level of a patient transfused with conventional stored blood.

[0072] The methods of this disclosure provide, and include, reducing BUN levels in trauma patients, comprising supplying trauma patients requiring transfusion therapy with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the BUN level is reduced by at least 5% compared to the BUN level of a patient transfused with conventional stored blood. In another embodiment, the BUN level is reduced by at least 10% compared to the BUN level of a patient transfused with conventional stored blood. In another embodiment, the BUN level is reduced by at least 20% compared to the BUN level of a patient transfused with conventional stored blood. In another embodiment, the BUN level is reduced by at least 30% compared to the BUN level of a patient transfused with conventional stored blood. In another embodiment, the BUN level is reduced by at least 40% compared to the BUN level of a patient transfused with conventional stored blood. In another embodiment, the BUN level is reduced by at least 50% compared to the BUN level of a patient transfused with conventional stored blood. In another embodiment, BUN levels are reduced by at least 60% compared to the BUN levels of patients transfused with conventionally stored blood. In yet another embodiment, BUN levels are reduced by at least 70% compared to the BUN levels of patients transfused with conventionally stored blood. In yet another embodiment, BUN levels are reduced by at least 80% compared to the BUN levels of patients transfused with conventionally stored blood. In a further embodiment, BUN levels are reduced by at least 90% compared to the BUN levels of patients transfused with conventionally stored blood. In a further embodiment, BUN levels are reduced by between 1-10%, 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-95% compared to the BUN levels of patients transfused with conventionally stored blood.

[0073] The method of this disclosure provides, and includes, reducing BUN levels in trauma patients by 1.5 to 10 times, comprising supplying trauma patients with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the BUN level is reduced by 2 to 3 times compared to the BUN level of a patient transfused with conventional stored blood. In another embodiment, the BUN level is reduced by 3 to 4 times. In another embodiment, the BUN level is reduced by 4 to 10 times compared to the BUN level of a patient transfused with conventional stored blood. In another embodiment, the BUN level is reduced by 6 to 9 times compared to the BUN level of a patient transfused with conventional stored blood. In a further embodiment, the BUN level is reduced by 2 to 5 times compared to the BUN level of a patient transfused with conventional stored blood. In another embodiment, the BUN level is reduced by 10 to 100 times compared to the BUN level of a patient transfused with conventional stored blood.

[0074] The method of this disclosure provides, and includes, reducing BUN levels in trauma patients by at least 1.5 times, comprising supplying trauma patients requiring transfusion therapy with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the BUN level is reduced by at least 2 times compared to the BUN level of a patient transfused with conventional stored blood. In another embodiment, the BUN level is reduced by at least 3 times compared to the BUN level of a patient transfused with conventional stored blood. In another embodiment, the BUN level is reduced by at least 4 times compared to the BUN level of a patient transfused with conventional stored blood. In another embodiment, the BUN level is reduced by at least 5 times compared to the BUN level of a patient transfused with conventional stored blood. In a further embodiment, the BUN level is reduced by at least 6 times. In another embodiment, the BUN level is reduced by at least 7 times compared to the BUN level of a patient transfused with conventional stored blood. In another embodiment, the BUN level is reduced by at least eight times compared to the BUN level of a patient who received a transfusion using conventionally stored blood. In another embodiment, the BUN level is reduced by at least nine times compared to the BUN level of a patient who received a transfusion using conventionally stored blood. In another embodiment, the BUN level is reduced by at least ten times compared to the BUN level of a patient who received a transfusion using conventionally stored blood.

[0075] The method of the present disclosure provides, and includes, a reduction of the percentage of CD45+ neutrophils in trauma patients, comprising supplying trauma patients with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the percentage of CD45+ neutrophils is reduced by at least 5% compared to the CD45+ neutrophil level of patients transfused with conventional stored blood. In another embodiment, the percentage of CD45+ neutrophils is reduced by at least 10% compared to the CD45+ neutrophil level of patients transfused with conventional stored blood. In another embodiment, the percentage of CD45+ neutrophils is reduced by at least 20% compared to the CD45+ neutrophil level of patients transfused with conventional stored blood. In another embodiment, the percentage of CD45+ neutrophils is reduced by at least 30% compared to the CD45+ neutrophil level of patients transfused with conventional stored blood. In another embodiment, the percentage of CD45+ neutrophils is reduced by at least 40% compared to the CD45+ neutrophil level of patients transfused with conventionally stored blood. In another embodiment, the percentage of CD45+ neutrophils is reduced by at least 50% compared to the CD45+ neutrophil level of patients transfused with conventionally stored blood. In another embodiment, the percentage of CD45+ neutrophils is reduced by at least 60% compared to the CD45+ neutrophil level of patients transfused with conventionally stored blood. In another embodiment, the percentage of CD45+ neutrophils is reduced by at least 70% compared to the CD45+ neutrophil level of patients transfused with conventionally stored blood. In yet another embodiment, the percentage of CD45+ neutrophils is reduced by at least 80% compared to the CD45+ neutrophil level of patients transfused with conventionally stored blood. In a further embodiment, the percentage of CD45+ neutrophils is reduced by at least 90% compared to the CD45+ neutrophil levels of patients transfused using conventionally stored blood.In a further embodiment, the percentage of CD45+ neutrophils is reduced by between 1–10%, 10–20%, 20–30%, 30–40%, 40–50%, 50–60%, 60–70%, 70–80%, 80–90%, or 90–95% compared to the CD45+ neutrophil levels of patients transfused with conventionally stored blood.

[0076] The present disclosure provides, and includes, a method for reducing the percentage of CD45+ neutrophils in trauma patients requiring transfusion therapy by 1.5 to 10 times, comprising supplying the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the percentage of CD45+ neutrophils is reduced by 2 to 3 times compared to the CD45+ neutrophil level of a patient transfused with conventional stored blood. In another embodiment, the percentage of CD45+ neutrophils is reduced by 3 to 4 times compared to the CD45+ neutrophil level of a patient transfused with conventional stored blood. In yet another embodiment, the percentage of CD45+ neutrophils is reduced by 4 to 10 times compared to the CD45+ neutrophil level of a patient transfused with conventional stored blood. In another embodiment, the percentage of CD45+ neutrophils is reduced by 6 to 9 times compared to the CD45+ neutrophil level of patients transfused with conventionally stored blood. In yet another embodiment, the percentage of CD45+ neutrophils is reduced by 2 to 5 times compared to the CD45+ neutrophil level of patients transfused with conventionally stored blood. In yet another embodiment, the percentage of CD45+ neutrophils is reduced by 10 to 50 times compared to the CD45+ neutrophil level of patients transfused with conventionally stored blood.

[0077] The method of this disclosure provides, and includes, reducing the percentage of CD45+ neutrophils in trauma patients requiring transfusion therapy by at least 1.5 times, comprising supplying the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the percentage of CD45+ neutrophils is reduced by at least 2 times compared to the CD45+ neutrophil level of a patient transfused with conventional stored blood. In another embodiment, the percentage of CD45+ neutrophils is reduced by at least 3 times compared to the CD45+ neutrophil level of a patient transfused with conventional stored blood. In yet another embodiment, the percentage of CD45+ neutrophils is reduced by at least 4 times compared to the CD45+ neutrophil level of a patient transfused with conventional stored blood. In another embodiment, the percentage of CD45+ neutrophils is reduced by at least 5 times compared to the CD45+ neutrophil level of patients transfused with conventionally stored blood. In yet another embodiment, the percentage of CD45+ neutrophils is reduced by at least 6 times compared to the CD45+ neutrophil level of patients transfused with conventionally stored blood. In yet another embodiment, the percentage of CD45+ neutrophils is reduced by at least 7 times compared to the CD45+ neutrophil level of patients transfused with conventionally stored blood. In yet another embodiment, the percentage of CD45+ neutrophils is reduced by at least 8 times compared to the CD45+ neutrophil level of patients transfused with conventionally stored blood. In yet another embodiment, the percentage of CD45+ neutrophils is reduced by at least 9 times compared to the CD45+ neutrophil level of patients transfused with conventionally stored blood. In another embodiment, the percentage of CD45+ neutrophils is reduced by at least 10 times compared to the CD45+ neutrophil levels of patients transfused using conventionally stored blood.

[0078] The methods of this disclosure provide, and include, reducing CXCL1 levels in trauma patients requiring transfusion therapy, comprising supplying the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the CXCL1 level is reduced by at least 5% compared to the CXCL1 level of a patient transfused with conventional stored blood. In another embodiment, the CXCL1 level is reduced by at least 10% compared to the CXCL1 level of a patient transfused with conventional stored blood. In another embodiment, the CXCL1 level is reduced by at least 20% compared to the CXCL1 level of a patient transfused with conventional stored blood. In another embodiment, the CXCL1 level is reduced by at least 30% compared to the CXCL1 level of a patient transfused with conventional stored blood. In another embodiment, the CXCL1 level is reduced by at least 40% compared to the CXCL1 level of a patient transfused with conventional stored blood. In another embodiment, the CXCL1 level is reduced by at least 50% compared to the CXCL1 level of patients transfused with conventionally stored blood. In yet another embodiment, the CXCL1 level is reduced by at least 60% compared to the CXCL1 level of patients transfused with conventionally stored blood. In yet another embodiment, the CXCL1 level is reduced by at least 70% compared to the CXCL1 level of patients transfused with conventionally stored blood. In yet another embodiment, the CXCL1 level is reduced by at least 80% compared to the CXCL1 level of patients transfused with conventionally stored blood. In yet another embodiment, the CXCL1 level is reduced by at least 90% compared to the CXCL1 level of patients transfused with conventionally stored blood. In a further embodiment, CXCL1 levels are reduced by between 1-10%, 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-95% compared to CXCL1 levels in patients transfused with conventionally stored blood.

[0079] The present disclosure provides, and includes, a method for reducing the CXCL1 level in trauma patients by 1.5 to 10 times, comprising supplying the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the CXCL1 level is reduced by 2 to 3 times compared to the CXCL1 level of a patient transfused with conventional stored blood. In another embodiment, the CXCL1 level is reduced by 3 to 4 times compared to the CXCL1 level of a patient transfused with conventional stored blood. In another embodiment, the CXCL1 level is reduced by 4 to 10 times compared to the CXCL1 level of a patient transfused with conventional stored blood. In yet another embodiment, the CXCL1 level is reduced by 6 to 9 times compared to the CXCL1 level of a patient transfused with conventional stored blood. In a further embodiment, the CXCL1 level is reduced by 2 to 5 times compared to the CXCL1 level of a patient transfused with conventional stored blood. In another embodiment, CXCL1 levels are reduced by 10 to 100 times compared to CXCL1 levels in patients transfused with conventionally stored blood.

[0080] The method of this disclosure provides, and includes, reducing the CXCL1 level in a trauma patient by at least 1.5 times, comprising supplying the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the CXCL1 level is reduced by at least 2 times compared to the CXCL1 level of a patient transfused with conventional stored blood. In another embodiment, the CXCL1 level is reduced by at least 3 times compared to the CXCL1 level of a patient transfused with conventional stored blood. In another embodiment, the CXCL1 level is reduced by at least 4 times compared to the CXCL1 level of a patient transfused with conventional stored blood. In another embodiment, the CXCL1 level is reduced by at least 5 times compared to the CXCL1 level of a patient transfused with conventional stored blood. In a further embodiment, the CXCL1 level is reduced by at least 6 times compared to the CXCL1 level of a patient transfused with conventional stored blood. In another embodiment, the CXCL1 level is reduced by at least seven times compared to the CXCL1 level of patients who received transfusions using conventionally stored blood. In another embodiment, the CXCL1 level is reduced by at least eight times compared to the CXCL1 level of patients who received transfusions using conventionally stored blood. In another embodiment, the CXCL1 level is reduced by at least nine times compared to the CXCL1 level of patients who received transfusions using conventionally stored blood. In another embodiment, the CXCL1 level is reduced by at least ten times compared to the CXCL1 level of patients who received transfusions using conventionally stored blood.

[0081] The present disclosure provides a method for reducing IL-6 levels in trauma patients requiring transfusion therapy, comprising supplying the trauma patient with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, the IL-6 level is reduced by at least 5% compared to the IL-6 level of a patient transfused with conventional stored blood. In another embodiment, the IL-6 level is reduced by at least 10% compared to the IL-6 level of a patient transfused with conventional stored blood. In another embodiment, the IL-6 level is reduced by at least 20%. In another embodiment, the IL-6 level is reduced by at least 30% compared to the IL-6 level of a patient transfused with conventional stored blood. In another embodiment, the IL-6 level is reduced by at least 40% compared to the IL-6 level of a patient transfused with conventional stored blood. In another embodiment, the IL-6 level is reduced by at least 50% compared to the IL-6 level of a patient transfused with conventional stored blood. In another embodiment, IL-6 levels are reduced by at least 60% compared to IL-6 levels in patients transfused with conventionally stored blood. In yet another embodiment, IL-6 levels are reduced by at least 70% compared to IL-6 levels in patients transfused with conventionally stored blood. In yet another embodiment, IL-6 levels are reduced by at least 80% compared to IL-6 levels in patients transfused with conventionally stored blood. In a further embodiment, IL-6 levels are reduced by at least 90% compared to IL-6 levels in patients transfused with conventionally stored blood. In a further embodiment, IL-6 levels are reduced by between 1-10%, 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-95% compared to IL-6 levels in patients transfused with conventionally stored blood.

[0082] The present disclosure provides, and includes, a method for reducing IL-6 levels in trauma patients by 1.5 to 10 times, comprising supplying trauma patients with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, IL-6 levels are reduced by 2 to 3 times compared to the IL-6 levels of patients transfused with conventional stored blood. In another embodiment, IL-6 levels are reduced by 3 to 4 times compared to the IL-6 levels of patients transfused with conventional stored blood. In yet another embodiment, IL-6 levels are reduced by 4 to 10 times. In yet another embodiment, IL-6 levels are reduced by 6 to 9 times compared to the IL-6 levels of patients transfused with conventional stored blood. In a further embodiment, IL-6 levels are reduced by 2 to 5 times compared to the IL-6 levels of patients transfused with conventional stored blood. In another embodiment, IL-6 levels are reduced by 10 to 100 times compared to IL-6 levels in patients transfused using conventionally stored blood.

[0083] The present disclosure provides, and includes, a method for reducing IL-6 levels in trauma patients by at least 1.5 times, comprising supplying trauma patients requiring transfusion therapy with oxygen-reduced stored blood having an oxygen saturation of 20% or less before and during storage. In one embodiment, IL-6 levels are reduced by at least 2 times compared to IL-6 levels in patients transfused with conventional stored blood. In another embodiment, IL-6 levels are reduced by at least 3 times compared to IL-6 levels in patients transfused with conventional stored blood. In another embodiment, IL-6 levels are reduced by at least 4 times compared to IL-6 levels in patients transfused with conventional stored blood. In another embodiment, IL-6 levels are reduced by at least 5 times. In a further embodiment, IL-6 levels are reduced by at least 6 times compared to IL-6 levels in patients transfused with conventional stored blood. In another embodiment, IL-6 levels are reduced by at least 7 times compared to IL-6 levels in patients transfused with conventional stored blood. In another embodiment, IL-6 levels are reduced by at least eightfold compared to IL-6 levels in patients who received transfusions using conventionally stored blood. In another embodiment, IL-6 levels are reduced by at least ninefold compared to IL-6 levels in patients who received transfusions using conventionally stored blood. In another embodiment, IL-6 levels are reduced by at least tenfold compared to IL-6 levels in patients who received transfusions using conventionally stored blood.

[0084] As used herein, the terms “higher,” “greater,” or “increased” mean that measurements of oxygen-reduced and anaerobic stored blood are at least one standard deviation greater for each measurement condition being compared to measurements of conventionally stored blood treated similarly by another method, with a sample size of two or more.

[0085] As used herein, the terms “reduced,” “lower,” “reduced,” or “less” mean that the measurements of oxygen-reduced and anaerobic stored blood are at least one standard deviation lower for each measurement condition being compared to measurements of conventionally stored blood RBCs treated similarly with normal oxygen or hyperoxygen, with a sample size of five or more samples.

[0086] As used herein, the term "approximately" refers to a range of ±10%.

[0087] As used herein, "less than" refers to a quantity that is smaller than zero.

[0088] The terms "comprises," "comprising," "includes," "including," and "having," as well as their conjugations, all mean to include but not be limited to what follows.

[0089] The phrase "consisting of" means "to include and limit what follows."

[0090] The term "essentially derived from" means that a composition, method, or structure may include additional components, steps, and / or parts, provided that such additional components, steps, and / or parts do not substantially alter the basic and novel properties of the claimed composition, method, or structure.

[0091] As used herein, the singular forms "a," "an," and "the" refer to multiple subjects unless otherwise clearly indicated by the context. For example, "compound" or "at least one compound" may include multiple compounds and mixtures thereof.

[0092] As used herein, the term “blood” refers to whole blood, leukocyte-reduced RBCs, platelet-reduced RBCs, and leukocyte and platelet-reduced RBCs. The term “blood” further includes concentrated red blood cells, platelet-reduced concentrated red blood cells, leukocyte-reduced concentrated red blood cells, and leukocyte and platelet-reduced concentrated red blood cells. The temperature of blood may vary depending on the stage of the collection process, starting at 37°C, the normal body temperature at the time of collection, but rapidly decreasing to about 30°C once outside the patient’s body, and then further decreasing to room temperature in about 6 hours if left untreated, and finally being refrigerated to between 4°C and 6°C. Human red blood cells in vivo are in a dynamic state. Red blood cells contain hemoglobin, an iron-containing protein that carries oxygen throughout the body and gives blood its red color. The percentage of blood volume composed of red blood cells is called hematocrit. As used herein, unless otherwise limited, RBC also includes concentrated red blood cells (pRBCs). Concentrated red blood cells are prepared from whole blood using centrifugation techniques commonly known in the art. As used herein, unless otherwise specified, the hematocrit of pRBCs is approximately 70%. As used herein, oxygen-reduced storage RBCs may include oxygen and carbon dioxide-reduced storage RBCs. As used herein, oxygen-reduced (OR) blood may include oxygen and carbon dioxide-reduced (OCR) blood.

[0093] As used herein, the terms “patient” and “subject” are interchangeable and refer to a human or animal in need of a blood transfusion.

[0094] As used herein, the term “trauma” includes blood loss and hemorrhagic trauma.

[0095] As used herein, the term “hemorrhagic shock” means shock resulting from loss of circulating blood volume and / or oxygen-carrying capacity. Hemorrhagic shock results from any condition associated with blood loss, internal bleeding (e.g., gastrointestinal bleeding) or external bleeding, and in particular trauma (e.g., penetrating or blunt trauma).

[0096] As used herein, the term “adverse event” includes events resulting from hemorrhagic shock in patients with hemorrhagic trauma.

[0097] As used herein, the terms “injury,” “damage,” and “failure” refer to organs in humans or animals that are not functioning properly or not functioning as expected, without disease or injury.

[0098] As used herein, a "unit" of blood is approximately 450-500 ml, including the anticoagulant. Suitable anticoagulants include CPD, CPDA1, ACD, and ACD-A.

[0099] Throughout this application, various aspects of the disclosure may be presented in the form of scope. It should be understood that scope descriptions are merely for convenience and brevity and should not be interpreted as inflexible limitations on the scope of the disclosure. Therefore, scope descriptions should be considered to specifically disclose all possible sub-scopes, as well as the individual numbers within that scope. For example, a scope description such as "1 to 6" should be considered to specifically disclose the individual numbers within that scope, such as 1, 2, 3, 4, 5, and 6, as well as sub-scopes such as "1 to 3," "1 to 4," "1 to 5," "2 to 4," "2 to 6," and "3 to 6." This applies regardless of the breadth of the scope.

[0100] Wherever a numerical range is indicated herein, it is meant to include any number (fractions and integers) listed within that range. The expressions "range between the first and second indicators" and "range from the first to the second indicator" are used interchangeably herein and are intended to include the first and second indicators as well as all fractions and integers between them.

[0101] As used herein, the term “method” means, means, techniques, and procedures for performing a given task, including, but not limited to, supplying a human patient in need of blood transfusion with oxygen-reduced stored blood having an initial oxygen saturation of 20% or less and stored for at least two days.

[0102] While this disclosure has been described with reference to specific embodiments, it will be understood by those skilled in the art that various modifications can be made without departing from the scope of this disclosure, and that equivalents can be used in place of elements of this disclosure. In addition, many modifications can be made without departing from the scope of this disclosure to adapt the teachings of this disclosure to specific circumstances or materials.

[0103] Therefore, this disclosure is not intended to be limited to any specific embodiment disclosed as the best mode contemplated for the implementation of this disclosure, but rather to include all embodiments that fall within the spirit and scope of the accompanying claims. [Examples]

[0104] Example 1: Blood collection and sample preparation Red blood cells are collected from a total of 12-14 rats in each pool in CP2D anticoagulant solution. The pooled blood is leukocyte-reduced using a neonatal leukocyte-reducing filter to separate its components, and the RBCs are stored in an AS-3 solution. RBCs are collected from a total of two pools. Each pool is divided into four types: untreated control (C), sham control (SC), oxygen-reduced (OR), and oxygen and carbon dioxide-reduced (OCR). For the C, SC, OR, and OCR units, the RBC subunits are processed by transferring them to 80 mL PVC transfusion bags, and the final RBC product is prepared by gas exchange. The RBC bags other than C are filled with 100% N2 (for OR), 95% N2 / 5% CO2 (for OCR), or air (SC) through a sterile filter, and gently rotated at 2-3 RPM along their long side (other than C). For OR and OCR units, the gas is removed through a filter after 10 minutes, and fresh gas is introduced for the subsequent gas exchange process. This process is repeated 5 to 8 times until the target SO2% of 5-10% is achieved as measured with an ABL-90 CO oximeter (Radiometer Copenhagen). SC units are rotated for 60 minutes without any gas exchange. OR and OCR units are stored anaerobically in canisters filled with N2, while C and SC units are stored in ambient air. All units are stored at 4°C for 3 weeks, and samples are taken on days 0 or 1, 7, 14, 21, and 28. Two pools were prepared and stored.

[0105] Analysis of ATP, 2,3-DPG, and hemolysis was performed on days 0, 1, 7, 14, 21, and 28. As shown in Figure 1, ATP levels were higher in OR blood on day 21 and in OCR blood on days 7, 14, 21, and 28 compared to conventional stored blood (control). OR blood also had higher levels of 2,3-DPG compared to the control on days 2, 7, and 14. OCR blood also showed higher levels of 2,3-DPG compared to the control on days 2, 7, and 14. See Figure 2.

[0106] Example 2: Recovery rate of oxygen-reduced blood Small amounts (less than 200 μL) of control, OR, and OCR blood stored for 3 weeks were labeled with technetium-99m. Animals were transfused with labeled RBCs (200 μL), and circulating radioactivity was periodically measured up to 24 hours after transfusion to estimate the percentage of viable transfused RBCs 24 hours after transfusion. As shown in Figure 3, when RBCs were stored for 3 weeks, OR- and OCR-RBCs were recovered in significantly greater numbers than control RBCs.

[0107] Example 3: A rat model of hemorrhagic shock resuscitation Blood collection and sample preparation: Red blood cells are collected from a total of 12-14 rats in CP2D anticoagulant solution from each pool. The pooled blood is leukocyte-reduced using a neonatal leukocyte-reducing filter to separate its components, and the RBCs are stored in an AS-3 solution. RBCs are collected from a total of six pools. Two pools are prepared for conventional storage (control). Two pools are oxygen-depleted (oxygen reduction; OR), and the remaining two blood pools are oxygen and carbon dioxide-depleted (oxygen and carbon dioxide reduction; OCR). Each of the four reduction pools is processed by transferring the RBCs to a 600 mL PVC transfusion bag, and the final RBC product is produced by gas exchange. The RBC bag is filled with 100% N2 (for OR) or 95% N2 / 5% CO2 (for OCR) through a sterile filter and gently rotated at 60-90 RPM along its long side. After 10 minutes, the gas is removed through a filter, and fresh gas is introduced for the subsequent gas exchange process. This process is repeated 5 to 8 times until the target SO2% of 5-10% is achieved, as measured by an ABL-90 CO oximeter (Radiometer Copenhagen). OR and OCR blood are stored anaerobically in canisters filled with N2.

[0108] The experiment will be conducted using Sprague-Dawley rats weighing 150-200 grams (g) (Charles River Laboratories, Boston, MA). Briefly, the animals will be anesthetized by intraperitoneal administration of 40 mg / kg of pentobarbital sodium. The animals will be placed supine on a heating pad to maintain a core body temperature of 37°C. The animals will be prepared for (i) catheter insertion into the left jugular vein and left femoral artery, (ii) tracheostomy (polyethylene-90 tubing), and (iii) introduction of a left ventricular (LV) conductance catheter via the right carotid artery. The animals will be subjected to mechanical ventilation using room air (TOPO ventilator (Kent Scientific, Torrington, CT)) at a respiratory rate of 50-70 breaths per minute and a supply of 10-15 cmH2O. After measurement, volatile anesthetic (1.5% / vol isoflurane (Dragerwerk AG, Laubeck, Germany)) was administered using a vaporizer connected to a ventilator. The depth of anesthesia was continuously monitored by pinching the toes, and the isoflurane was increased by 0.1% / vol increments as needed to prevent discomfort to the animals.

[0109] Anesthetized animals are subjected to hypovolemic shock by inducing bleeding by removing 50% of their blood volume via a femoral artery catheter within 10 minutes. This hypovolemic shock state is maintained for 30 minutes. Resuscitation is performed by infusing pre-stored red blood cells (RBCs) via the femoral artery at a rate of 300 microliters per minute (μL / min) over a 60-minute period until the mean arterial pressure (MAP) stabilizes at 90% of baseline. MAP and heart rate (HR) are acquired via the femoral artery catheter at 10, 20, 30, 45, and 60 minutes during this period (using a PowerLab (AD Instruments, Colorado Springs, CO)). After 60 minutes, hematocrit (Hct) is measured by centrifugation of heparinized capillaries. Hemoglobin (Hb), lactate, glucose, K+, Na+, pH, and arterial blood gases are quantified using an ABL90 CO oximeter (Radiometer, Copenhagen). Cardiac function indices and systemic values ​​(MAP, HR, Hct, Hb, and blood gases) are monitored at baseline (BL), during shock, and at 10 minutes (early resuscitation), 20 minutes, 30 minutes, 45 minutes, and 60 minutes (late resuscitation) after resuscitation. Animals are euthanized at the end of the experiment.

[0110] Example 4: Hematocrit analysis in a rat model of hemorrhagic shock Hematocrit (Hct) decreases by approximately 30–40% after hypovolemic shock is induced. Hematocrit can be restored to normal levels by supplying conventional, OR, or OCR blood stored for one week. See Figure 4A. However, as shown in Figure 4B, OR blood stored for one week shows an increased hematocrit percentage compared to control and OCR blood at 10 minutes after resuscitation (early R). The hematocrit percentage of OR blood remains improved compared to control at 60 minutes after resuscitation (late R).

[0111] Example 5: Changes in mean arterial pressure due to oxygen-reduced blood Mean arterial pressure (MAP) is obtained from a femoral artery catheter (PowerLab (AD Instruments, Colorado Springs, CO)). As shown in Figure 5A, baseline MAP is between 80 and 110 mmHg. MAP decreases to between 20 and 60 mmHg during hemorrhagic shock. Resuscitation of animals using OR and OCR blood stored for one week increases MAP to approximately 80 and 90 mmHg, respectively. As shown in Figure 5B, resuscitation with OR blood can restore MAP to the normal range after 10 minutes compared to the control. Control and OCR stored blood can restore MAP to the normal range after 60 minutes of resuscitation. The amount of blood required for resuscitation and hemodynamic maintenance using conventional stored RBCs (control) was greater than the amount of OR and OCR RBCs required. See Figures 6A and 6B.

[0112] Example 6: Metabolic response to hemorrhagic shock Hemorrhagic shock in animals increases lactate levels from approximately 2 mmol / L to approximately 8–14 mmol / L. Resuscitation using OR and OCR RBCs stored for one week reduces lactate to near normal levels exactly 10 minutes after resuscitation. See Figure 7A. Lactate levels in animals resuscitated with control blood were similar to those in animals under hemorrhagic shock. Animals treated for 60 minutes with control, OR, and OCR RBCs showed similar lactate levels. As shown in Figure 7B, OR RBCs stored for three weeks could also reduce lactate levels compared to the control 10 minutes after resuscitation. However, after 80 minutes after resuscitation, OCR RBCs restored lactate levels to the normal range. While control and OR RBCs were able to reduce lactate levels, they could not reduce them to the normal range of 1–3 mmol / L. Analysis of glucose levels showed that in animals under hemorrhagic shock, glucose levels increased from the normal range of approximately 160 mg / dL to 240 mg / dL to approximately 320 to 510 mg / dL. See Figures 8A and 8B. Both OR and OCR RBCs stored for one week showed a decrease in glucose levels compared to the control 10 minutes after resuscitation. All three samples recovered glucose levels to the normal range 60 minutes after resuscitation. As shown in Figure 8B, OR and OR RBCs stored for three weeks also showed a decrease in glucose levels compared to the control 10 minutes after resuscitation. Unlike RBCs stored for one week, only OR and OCR RBCs were able to recover glucose to the normal range. Therefore, both lactate and glucose levels were reduced more rapidly in OR and OCR RBCs compared to control RBCs.

[0113] Example 7: Injury and inflammation of vital organs Organ injury and inflammation were analyzed in animals after hemorrhagic shock and resuscitation. Elevated levels of liver enzymes indicate some form of liver injury or damage. Liver injury was quantified by analyzing aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels. Resuscitation using OR and OCR RBCs stored for 1 week prior (Figures 9A and 10A) and OR and OCR RBCs stored for 3 weeks prior (Figures 9B and 10B) reduced AST and ALT levels compared to control RBCs stored for the same period. Renal function was quantified by analyzing serum creatinine and blood urea nitrogen (BUN) levels. OR and OCR RBCs stored for 1 week reduced serum creatinine levels by more than 30% compared to control RBCs (Figure 11A). After 3 weeks of storage, serum creatinine levels in animals treated with control, OR, and OCR RBCs overlapped (Figure 11B). BUN levels were more than 30% lower in animals treated with OCR RBCs stored for one week compared to the control group (Figure 12A). Similarly, BUN levels were more than 30% lower in animals treated with OR RBCs stored for three weeks compared to the control group (Figure 12B). Overall, vital organ function was preserved more effectively with OR and OCR RBCs compared to control RBCs.

[0114] At the completion of the in vivo study, the liver, spleen, and lungs were removed from the animals and analyzed for various inflammatory factors, including CXCL1, urinary neutrophil gelatinase-associated lipocalin (u-NGAL), IL-6, and neutrophils. CXCL1 levels were reduced in animals treated with OR and OCR RBCs stored for 1 or 3 weeks compared to controls stored for the same period (Figures 13A, B, 14A, B, and 15A, B). As shown in Figures 16A and B, u-NGAL levels were reduced in the kidneys of animals treated with OR and OCR RBCs stored for 1 or 3 weeks compared to control RBCs stored for a comparable period (Figure 16). As shown in Figures 17 and 18, the percentage levels of CD45+ neutrophils and IL-6 in the lungs removed from the animals were significantly lower in OR and OCR RBCs compared to control RBCs stored for the same period. These results indicate that organ damage and inflammation were reduced in animals treated with OR and OCR RBCs compared to animals treated with control RBCs.

Claims

1. Oxygen- and carbon dioxide-reduced stored blood for use in reducing the risk of organ damage in patients with hemorrhagic trauma where reducing the risk of organ damage is necessary, The oxygen and carbon dioxide-reduced stored blood has an initial oxygen saturation (SO4) of 10% or less. 2 ) It has a level of 10% or less SO2 during an anaerobic storage period of up to 3 weeks. 2 Maintain the level, The oxygen- and carbon dioxide-reduced preserved blood is used in patients with hemorrhagic trauma for whom the risk of organ damage needs to be reduced, and which has an elevated lactate level of 8 to 14 mmol / L (mole / L) before administration of the oxygen- and carbon dioxide-reduced preserved blood.

2. The oxygen and carbon dioxide reduced stored blood according to claim 1, wherein the organ damage is selected from the group consisting of liver injury, lung failure, kidney failure, and heart failure.

3. The oxygen and carbon dioxide reduced stored blood according to claim 1, wherein the patient with hemorrhagic trauma requiring reduction of the risk of organ damage has elevated levels of urinary neutrophil gelatinase-related lipocalin (u-NGAL) before administration.

4. The oxygen and carbon dioxide reduced stored blood according to claim 1, wherein the patient with hemorrhagic trauma requiring a reduction in the risk of organ damage has a reduced hematocrit before administration.

5. The oxygen and carbon dioxide reduced stored blood according to claim 1, wherein the elevated lactate level in hemorrhagic trauma patients requiring reduction of the risk of organ damage is reduced by at least 10% after administration compared to hemorrhagic trauma patients administered conventional stored blood that does not have reduced oxygen and carbon dioxide.

6. The oxygen and carbon dioxide reduced stored blood according to claim 3, wherein the elevated u-NGAL level in hemorrhagic trauma subjects requiring reduction of the risk of organ damage is reduced by at least 10% after administration compared to hemorrhagic trauma subjects administered conventional stored blood that does not have reduced oxygen and carbon dioxide.

7. The oxygen- and carbon dioxide-reduced stored blood according to claim 4, wherein the reduced hematocrit in hemorrhagic trauma patients requiring a reduction in the risk of organ damage increases after administration compared to hemorrhagic trauma patients administered conventional stored blood that does not have reduced oxygen and carbon dioxide.

8. Initial oxygen saturation (SO4) is used to reduce elevated lactate levels in patients with hemorrhagic trauma who require a reduction in elevated lactate levels. 2 ) Levels below 10%, and during anaerobic storage periods of up to 3 weeks, SO levels below 10% 2 Blood stored with reduced oxygen and carbon dioxide levels, The aforementioned use includes administering oxygen- and carbon dioxide-reduced stored blood to a patient with hemorrhagic trauma who requires a reduction in elevated lactate levels. The blood of a patient with hemorrhagic trauma requiring a reduction in elevated lactate levels has elevated lactate levels of 8-14 mmol / L (mole / L) before administration, and these lactate levels indicate hemorrhagic shock before administration, and The oxygen- and carbon dioxide-reduced stored blood in which the hemorrhagic shock recovers after administration.

9. The oxygen and carbon dioxide reduced stored blood according to claim 8, wherein the hemorrhagic trauma patient requiring a reduction in elevated lactate levels is at risk of developing organ damage selected from the group consisting of liver injury, lung failure, kidney failure, and heart failure.

10. The oxygen and carbon dioxide reduced blood storage method according to claim 1 or 8, wherein the hemorrhagic trauma patient requiring a reduction in elevated lactate levels further exhibits elevated levels of one or more liver enzymes selected from the group consisting of aspartate aminotransferase (AST), alanine aminotransferase (ALT), and combinations thereof, prior to administration.

11. The oxygen and carbon dioxide reduced stored blood according to any one of claims 1 to 10, wherein the patient with hemorrhagic trauma requiring a reduction in elevated lactate levels has a glucose level exceeding 200 milligrams / deciliter (mg / dL) before administration.

12. The oxygen and carbon dioxide reduced stored blood according to any one of claims 1 to 11, wherein the patient with hemorrhagic trauma requiring a reduction in elevated lactate levels exhibits a low mean arterial pressure before administration.

13. The oxygen and carbon dioxide reduced stored blood according to any one of claims 1 to 12, wherein the patient with hemorrhagic trauma requiring a reduction in elevated lactate levels exhibits elevated values ​​of one or more renal enzymes selected from the group consisting of serum creatinine and blood urea nitrogen (BUN) prior to administration.

14. The oxygen and carbon dioxide reduced blood storage method according to claim 8, wherein the patient with hemorrhagic trauma requiring a reduction in elevated lactate levels exhibits elevated levels of one or more inflammatory factors selected from the group consisting of CXC motif chemokine ligand 1 (CXCL1), urinary neutrophil gelatinase-related lipocalin (u-NGAL), interleukin-6 (IL-6), and neutrophils prior to administration.

15. The oxygen and carbon dioxide reduced stored blood according to claim 8, wherein the hemorrhagic trauma subject requiring a reduction in elevated lactate levels is selected from the group consisting of surgery, penetrating wounds, blunt force trauma, injuries from falls, and injuries from traffic accidents.

16. The oxygen and carbon dioxide-reduced stored blood according to claim 8, wherein the elevated lactate level decreases by 10-90% after administration.

17. The oxygen and carbon dioxide-reduced stored blood according to claim 8, wherein the elevated lactate level is reduced by at least 20% after administration.

18. The oxygen- and carbon dioxide-reduced stored blood according to claim 12, wherein the low mean arterial pressure of hemorrhagic trauma patients requiring a reduction in elevated lactate levels is elevated after administration compared to hemorrhagic trauma patients administered conventional stored blood that has not had its oxygen and carbon dioxide levels reduced.

19. The oxygen- and carbon dioxide-reduced stored blood according to claim 10, wherein the increase in the one or more liver enzymes is reduced by at least 5% after administration compared to hemorrhagic trauma subjects who were administered conventional stored blood in which oxygen and carbon dioxide levels were not reduced.

20. The oxygen- and carbon dioxide-reduced stored blood according to claim 14, wherein the increased u-NGAL level is reduced by at least 5% after administration compared to hemorrhagic trauma subjects who were administered conventional stored blood that has not had its oxygen and carbon dioxide reduced.

21. The oxygen- and carbon dioxide-reduced stored blood according to claim 13, wherein the increase in the one or more renal enzymes is at least 5% lower after administration compared to hemorrhagic trauma subjects who were administered conventional stored blood that has not had its oxygen and carbon dioxide reduced.

22. The oxygen- and carbon dioxide-reduced stored blood according to claim 11, wherein the glucose level is at least 10% lower after administration compared to hemorrhagic trauma subjects who were administered conventional stored blood that has not had its oxygen and carbon dioxide reduced.

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